The present application claims benefit of Japanese Patent Application Nos. Hei. 11-377484 (filed Dec. 16, 1999), 2000-159162 (filed Apr. 7, 2000) and 2000-280988 (filed Aug. 3, 2000), the entire contents of each of which is incorporated herein by reference.
The contents of the attached CD-R compact discs are incorporated herein by reference in their entirety. The attached discs contain an identical copy of a file “SEQ2.TXT” which were created on the discs on Dec. 13, 2000, and are each 25,891 KB.
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to novel polynucleotides derived from microorganisms belonging to coryneform bacteria and fragments thereof, polypeptides encoded by the polynucleotides and fragments thereof, polynucleotide arrays comprising the polynucleotides and fragments thereof, computer readable recording media in which the nucteotide sequences of the polynucleotide and fragments thereof have been recorded, and use of them as well as a method of using the polynucleotide and/or polypeptide sequence information to make comparisons.
2. Brief Description of the Background Art
Coryneform bacteria are used in producing various useful substances, such as amino acids, nucleic acids, vitamins, saccharides (for example, ribulose), organic acids (for example, pyruvic acid), and analogues of the above-described substances (for example, N-acetylamino acids) and are very useful microorganisms industrially. Many mutants thereof are known.
For example, Corynebacterium glutamicum is a Gram-positive bacterium identified as a glutamic acid-producing bacterium, and many amino acids are produced by mutants thereof. For example, 1,000,000 ton/year of L-glutamic acid which is useful as a seasoning for umami (delicious taste), 250,000 ton/year of L-lysine which is a valuable additive for livestock feeds and the like, and several hundred ton/year or more of other amino acids, such as L-arginine, L-proline, L-glutamine, L-tryptophan, and the like, have been produced in the world (Nikkei Bio Yearbook 99, published by Nikkei BP (1998)).
The production of amino acids by Corynebacterium glutamicum is mainly carried out by its mutants (metabolic mutants) which have a mutated metabolic pathway and regulatory systems. In general, an organism is provided with various metabolic regulatory systems so as not to produce more amino acids than it needs. In the biosynthesis of L-lysine, for example, a microorganism belonging to the genus Corynebacterium is under such regulation as preventing the excessive production by concerted inhibition by lysine and threonine against the activity of a biosynthesis enzyme common to lysine, threonine and methionine, i.e., an aspartokinase, (J. Biochem., 65: 849-859 (1969)). The biosynthesis of arginine is controlled by, repressing the expression of its biosynthesis gene by arginine so as not to biosynthesize an excessive amount of arginine (Microbiology, 142: 99-108 (1996)). It is considered that these metabolic regulatory mechanisms are deregulated in amino acid-producing mutants. Similarly, the metabolic regulation is deregulated in mutants producing nucleic acids, vitamins, saccharides, organic acids and analogues of the above-described substances so as to improve the productivity of the objective product.
However, accumulation of basic genetic, biochemical and molecular biological data on coryneform bacteria is insufficient in comparison with Escherichia coli, Bacillus subtilis, and the like. Also, few findings have been obtained on mutated genes in amino acid-producing mutants. Thus, there are various mechanisms, which are still unknown, of regulating the growth and metabolism of these microorganisms.
A chromosomal physical map of Corynebacterium glutamicum ATCC 13032 is reported and it is known that its genome size is about 3,100 kb (Mol. Gen. Genet., 252: 255-265 (1996)). Calculating on the basis of the usual gene density of bacteria, it is presumed that about 3,000 genes are present in this genome of about 3,100 kb. However, only about 100 genes mainly concerning amino acid biosynthesis genes are known in Corynebacterium glutamicum, and the nucleotide sequences of most genes have not been clarified hitherto.
In recent years, the full nucleotide sequence of the genomes of several microorganisms, such as Escherichia coli, Mycobacterium tuberculosis, yeast, and the like, have been determined (Science, 277: 1453-62 (1997); Nature, 393: 537-544 (1998); Nature, 387: 5-105 (1997)). Based on the thus determined full nucleotide sequences, assumption of gene regions and prediction of their function by comparison with the nucleotide sequences of known genes have been carried out. Thus, the functions of a great number of genes have been presumed, without genetic, biochemical or molecular biological experiments.
In recent years, moreover, techniques for monitoring expression levels of a great number of genes simultaneously or detecting mutations, using DNA chips, DNA arrays or the like in which a partial nucleic acid fragment of a gene or a partial nucleic acid fragment in genomic DNA other than a gene is fixed to a solid support, have been developed. The techniques contribute to the analysis of microorganisms, such as yeasts, Mycobacterium tuberculosis, Mycobacterium bovis used in BCG vaccines, and the like (Science, 278: 680-686 (1997); Proc. Natl. Acad. Sci. USA, 96: 12833-38 (1999); Science, 284: 1520-23 (1999)).
SUMMARY OF THE INVENTION
An object of the present invention is to provide a polynucleotide and a polypeptide derived from a microorganism of coryneform bacteria which are industrially useful, sequence information of the polynucleotide and the polypeptide, a method for analyzing the microorganism, an apparatus and a system for use in the analysis, and a method for breeding the microorganism.
The present invention provides a polynucleotide and an oligonucleotide derived from a microorganism belonging to coryneform bacteria, oligonucleotide arrays to which the polynucleotides and the oligonucleotides are fixed, a polypeptide encoded by the polynucleotide, an antibody which recognizes the polypeptide, polypeptide arrays to which the polypeptides or the antibodies are fixed, a computer readable recording medium in which the nucleotide sequences of the polynucleotide and the oligonucleotide and the amino acid sequence of the polypeptide have been recorded, and a system based on the computer using the recording medium as well as a method of using the polynucleotide and/or polypeptide sequence information to make comparisons.
BRIEF DESCRIPTION OF THE DRAWING
FIG. 1 is a map showing the positions of typical genes on the genome of Corynebacterium glutamicum ATCC 13032.
FIG. 2 is electrophoresis showing the results of proteome analyses using proteins derived from (A) Corynebacterium glutamicum ATCC 13032, (B) FERM BP-7134, and (C) FERM BP-158.
FIG. 3 is a flow chart of an example of a system using the computer readable media according to the present invention.
FIG. 4 is a flow chart of an example of a system using the computer readable media according to the present invention.
DETAILED DESCRIPTION OF THE INVENTION
This application is based on Japanese applications No. Hei. 11-377484 filed on Dec. 16, 1999, No. 2000-159162 filed on Apr. 7, 2000 and No. 2000-280988 filed on Aug. 3, 2000, the entire contents of which are incorporated hereinto by reference.
From the viewpoint that the determination of the full nucleotide sequence of Corynebacterium glutamicum would make it possible to specify gene regions which had not been previously identified, to determine the function of an unknown gene derived from the microorganism through comparison with nucleotide sequences of known genes and amino acid sequences of known genes, and to obtain a useful mutant based on the presumption of the metabolic regulatory mechanism of a useful product by the microorganism, the inventors conducted intensive studies and, as a result, found that the complete genome sequence of Corynebacterium glutamicum can be determined by applying the whole genome shotgun method.
Specifically, the present invention relates to the following (1) to (65):
(1) A method for at least one of the following:
(A) identifying a mutation point of a gene derived from a mutant of a coryneform bacterium,
(B) measuring an expression amount of a gene derived from a coryneform bacterium,
(C) analyzing an expression profile of a gene derived from a coryneform bacterium,
(D) analyzing expression patterns of genes derived from a coryneform bacterium, or
(E) identifying a gene homologous to a gene derived from a coryneform bacterium,
said method comprising:
(a) producing a polynucleotide array by adhering to a solid support at least two polynucleotides selected from the group consisting of first polynucleotides comprising the nucleotide sequence represented by any one of SEQ ID NOS:1 to 3501, second polynucleotides which hybridize with the first polynucleotides under stringent conditions, and third polynucleotides comprising a sequence of 10 to 200 continuous bases of the first or second polynucleotides,
(b) incubating the polynucleotide array with at least one of a labeled polynucleotide derived from a coryneform bacterium, a labeled polynucleotide derived from a mutant of the coryneform bacterium or a labeled polynucleotide to be examined, under hybridization conditions,
(c) detecting any hybridization, and
(d) analyzing the result of the hybridization.
As used herein, for example, the at least two polynucleotides can be at least two of the first polynucleotides, at least two of the second polynucleotides, at least two of the third polynucleotides, or at least two of the first, second and third polynucleotides.
(2) The method according to (1), wherein the coryneform bacterium is a microorganism belonging to the genus Corynebacterium, the genus Brevibacterium, or the genus Microbacterium.
(3) The method according to (2), wherein the microorganism belonging to the genus Corynebacterium is selected from the group consisting of Corynebacterium glutamicum, Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium callunae, Corynebacterium herculis, Corynebacterium lilium, Corynebacterium melassecola, Corynebacterium thermoaminogenes, and Corynebacterium ammoniagenes.
(4) The method according to (1), wherein the polynucleotide derived from a coryneform bacterium, the polynucelotide derived from a mutant of the coryneform bacterium or the polynucleotide to be examined is a gene relating to the biosynthesis of at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogues thereof.
(5) The method according to (1), wherein the polynucleotide to be examined is derived from Escherichia coli.
(6) A polynucleotide array, comprising:
at least two polynucleotides selected from the group consisting of first polynucleotides comprising the nucleotide sequence represented by any one of SEQ ID NOS:1 to 3501, second polynucleotides which hybridize with the first polynucleotides under stringent conditions, and third polynucleotides comprising 10 to 200 continuous bases of the first or second polynucleotides, and
a solid support adhered thereto.
As used herein, for example, the at least two polynucleotides can be at least two of the first polynucleotides, at least two of the second polynucleotides, at least two of the third polynucleotides, or at least two of the first, second and third polynucleotides.
(7) A polynucleotide comprising the nucleotide sequence represented by SEQ ID NO:1 or a polynucleotide having a homology of at least 80% with the polynucleotide.
(8) A polynucleotide comprising any one of the nucleotide sequences represented by SEQ ID NOS:2 to 3431, or a polynucleotide which hybridizes with the polynucleotide under stringent conditions.
(9) A polynucleotide encoding a polypeptide having any one of the amino acid sequences represented by SEQ ID NOS:3502 to 6931, or a polynucleotide which hybridizes therewith under stringent conditions.
(10) A polynucleotide which is present in the 5′ upstream or 3′ downstream of a polynucleotide comprising the nucleotide sequence of any one of SEQ ID NOS:2 on 3431 in a whole polynucleotide comprising the nucleotide sequence represented by SEQ ID NO:1, and has an activity of regulating an expression of the polynucleotide.
(11) A polynucleotide comprising 10 to 200 continuous bases in the nucleotide sequence of the polynucleotide of any one of (7) to (10), or a polynucleotide comprising a nucleotide sequence complementary to tie polynucleotide comprising 10 to 200 continuous based.
(12) A recombinant DNA comprising the polynucleotide of any one of (8) to (11).
(13) A transformant comprising the polynucleotide of any one of (8) to (11) or the recombinant DNA of (12).
(14) A method for producing a polypeptide, comprising:
culturing the transformant of (13) in a medium to produce and accumulate a polypeptide encoded by the polynucleotide of (8) or (9) in the medium, and
recovering the polypeptide from the medium.
(15) A method for producing at least one of an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogues thereof, comprising:
culturing the transformant of (13) in a medium to produce and accumulate at least one of an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogues thereof in the medium, and
recovering the at least one of the amino acid, the nucleic acid, the vitamin, the saccharide, the organic acid, and analogues thereof from the medium.
(16) A polypeptide encoded by a polynucleotide comprising the nucleotide sequence selected from SEQ ID NOS:2 to 3431.
(17) A polypeptide comprising the amino acid sequence selected from SEQ ID NOS:3502 to 6931.
(18) The polypeptide according to (16) or (17), wherein at least one amino acid is deleted, replaced, inserted or added, said polypeptides having an activity which is substantially the same as that of the polypeptide without said at least one amino acid deletion, replacement, insertion or addition.
(19) A polypeptide comprising an amino acid sequence having a homology of at least 60% with the amino acid sequence of the polypeptide of (16) or (17), and having an activity which is substantially the same as that of the polypeptide.
(20) An antibody which recognizes the polypeptide of any one of (16) to (19).
(21) A polypeptide array, comprising:
at least one polypeptide or partial fragment polypeptide selected from the polypeptides of (16) to (19) and partial fragment polypeptides of the polypeptides, and a solid support adhered thereto.
(22) A polypeptide array, comprising:
at least one antibody which recognizes a polypeptide or partial fragment polypeptide selected from the polypeptides of (16) to (19) and partial fragment polypeptides of the polypeptides, and
a solid support adhered thereto.
(23) A system based on a computer for identifying a target sequence or a target structure motif derived from a coryneform bacterium, comprising the following:
(i) a user input device that inputs at least one nucleotide sequence information selected from SEQ ID NOS:1 to 3501, and target sequence or target structure motif information;
(ii) a data storage device for at least temporarily storing the input information;
(iii) a comparator that compares the at least one nucleotide sequence information selected from SEQ ID NOS:1 to 3501 with the target sequence or target structure motif information, recorded by the data storage device for screening and analyzing nucleotide sequence information which is coincident with or analogous to the target sequence or target structure motif information; and
(iv) an output device that shows a screening or analyzing result obtained by the comparator.
(24) A method based on a computer for identifying a target sequence or a target structure motif derived from a coryneform bacterium, comprising the following:
(i) inputting at least one nucleotide sequence, information selected from SEQ ID NOS:1 to 3501, target sequence information or target structure motif information into a user input device;
(ii) at least temporarily storing said information;
(iii) comparing the at least one nucleotide sequence information selected from SEQ ID NOS:1 to 3501 with the target sequence or target structure motif information; and
(iv) screening and analyzing nucleotide sequence information which is coincident with or analogous to the target sequence or target structure motif information.
(25) A system based on a computer for identifying a target sequence or a target structure motif derived from a coryneform bacterium, comprising the following:
(i) a user input device that inputs at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001, and target sequence or target structure motif information;
(ii) a data storage device for at least temporarily storing the input information;
(iii) a comparator that compares the at least one amine acid sequence information selected from SEQ ID NOS:3502 to 7001 with the target sequence or target structure motif information, recorded by the data storage device for screening and analyzing amino acid sequence information which is coincident with or analogous to the target sequence or target structure motif information; and
(iv) an output device that shows a screening or analyzing result obtained by the comparator.
(26) A method based on a computer for identifying a target sequence or a target structure motif derived from a coryneform bacterium, comprising the following:
(i) inputting at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001, and target sequence information or target structure motif information into a user input device;
(ii) at least temporarily storing said information;
(iii) comparing the at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001 with the target sequence or target structure motif information; and
(iv) screening and analyzing amino acid sequence information which is coincident with or analogous to the target sequence or target structure motif information.
(27) A system based on a computer for determining a function of a polypeptide encoded by a polynucleotide having a target nucleotide sequence derived from a coryneform bacterium, comprising the following:
(i) a user input device that inputs at least one nucleotide sequence information selected from SEQ ID NOS:2 to 3501, function information of a polypeptide encoded by the nucleotide sequence, and target nucleotide sequence information;
(ii) a data storage device for at least temporarily storing the input information;
(iii) a comparator that compares the at least one nucleotide sequence information selected from SEQ ID NOS:2 to 3501 with the target nucleotide sequence information, and determining a function of a polypeptide encoded by a polynucleotide having the target nucleotide sequence which is coincident with or analogous to the polynucleotide having at least one nucleotide sequence selected from SEQ ID NOS:2 to 3501; and
(iv) an output devices that shows a function obtained by the comparator.
(28) A method based on a computer for determining a function of a polypeptide encoded by a polypeptide encoded by a polynucleotide having a target nucleotide sequence derived from a coryneform bacterium, comprising the following:
(i) inputting at least one nucleotide sequence information selected from SEQ ID NOS:2 to 3501, function information of a polypeptide encoded by the nucleotide sequence, and target nucleotide sequence information;
(ii) at least temporarily storing said information;
(iii) comparing the at least one nucleotide sequence information selected from SEQ ID NOS:2 to 3501 with the target nucleotide sequence information; and
(iv) determining a function of a polypeptide encoded by a polynucleotide having the target nucleotide sequence which is coincident with or analogous to the polynucleotide having at least one nucleotide sequence selected from SEQ ID NOS:2 to 3501.
(29) A system based on a computer for determining a function of a polypeptide having a target amino acid sequence derived from a coryneform bacterium, comprising the following:
(i) a user input device that inputs at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001, function information based on the amino acid sequence, and target amino acid sequence information;
(ii) a data storing device for at least temporarily storing the input information;
(iii) a comparator that compares the at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001 with the target amino acid sequence information for determining a function of a polypeptide having the target amino acid sequence which is coincident with or analogous to the polypeptide having at least one amino acid sequence selected from SEQ ID NOS: 3502 to 7001; and
(iv) an output device that shows a function obtained by the comparator.
(30) A method based on a computer for determining a function of a polypeptide having a target amino acid sequence derived from a coryneform bacterium, comprising the following:
(i) inputting at least one amino acid sequence information selected from SEQ ID NOS: 3502 to 7001, function information based on the amino acid sequence, and target amino acid sequence information;
(ii) at least temporarily storing said information;
(iii) comparing the at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001 with the target amino acid sequence information; and
(iv) determining a function of a polypeptide having the target amino acid sequence which is coincident with or analogous to the polypeptide having at least one amino acid sequence selected from SEQ ID NOS:3502 to 7001.
(31) The system according to any one of (23), (25), (27) and (29), wherein a coryneform bacterium is a microorganism of the genus Corynebacterium, the genus Brevibacterium, or the genus Microbacterinum
(32) The method according to any one of (24), (26), (28) and (30), wherein a coryneform bacterium is a microorganism of the genus Corynebacterium, the genus Brevibacterium , or the genus Microbacterinum.
(33) The system according to (31), wherein the microorganism belonging to the genus Corynebacterium is selected from the group consisting of Corynebacterium glutamicum, Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium callunae, Corynebacterium herculis, Corynebacterium lilium, Corynebacterium melassecola, Corynebacterium thermoaminogenes, and Corynebacterium ammoniagenes.
(34) The method according to (32), wherein the microorganism belonging to the genus Corynebacterium is selected from the grow consisting of Corynebacterium glutamicum, Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium callunae, Corynebacterium herculis, Corynebacterium lilium, Corynebacterium melassecola, Corynebacterium thermoaminogenes, and Corynebacterium ammoniagenes.
(35) A recording medium or storage device which is readable by a computer is which at least one nucleotide sequence information or selected from SEQ ID NOS:1 to 3501 or function information based or the nucleotide sequence is recorded, and is usuable in the system of (23) or (27) or the method of (24) or (28)
(36) A recording medium or storage device which is readable by a computer in which at least one amino acid sequence information selected from SEQ ID NOS:3502 to 7001 or function information based on the amino acid sequence is recorded, and is usable in the system of (25) or (29) or the method of (26) or (30).
(37) The recording medium or storage device according to (35) or (36), wherein is a computer readable recording medium selected from the group consisting of a floppy disc, a hard disc, a magnetic tape, a random access memory (RAM), a read only memory (ROM), a magneto-optic disc (MO), CD-ROM, CD-R, CD-RW, DVD-ROM, DVD-RAM and DVD-RW.
(38) A polypepide having a homoserine dehydrogenase activity, comprising an amino acid sequence in which the Val residue at the sequence in the amino acid sequence of homoserine dehydrogenase derived from a coryneform bacterium is replaced with amino acid residue other than a Val residue.
(39) A polypeptide comprising an amino acid sequence in which the Val residue at the 59th position at the amino acid sequence as represented by SEQ ID NO:6952 is replaced with amino acid residue other than a Val residue.
(40) The polypeptide according to (38) or (39) wherein the Val residue at the 59th position is replaced with an Ala residue.
(41) A polypeptide having private carboxylase activity, comprising an amino acid sequence in which the Pro residue at the 458th position in the amino acid sequence of private carboxylase derived from a coryneform bacterium is replaced with an amino acid residue other than a Pro residue.
(42) A polypeptide cruising an amino acid sequence in which the Pro residue at the 458th position in the amino acid sequence represented by SEQ ID NO:4265 is replaced with an amino acid residue other than a Pro residue.
(43) The polypeptide according to (41) or (42), wherein the Pro residue at the 458th position is replaced with a Ser residue.
(44) The polypeptide recording to any one of (38) to (43), which is derived from Corynebacterium glutamicum.
(45) A DNA encoding the polypeptide of any one of (38) to (44).
(46) A recombinant DNA comprising the DNA of (45).
(47) A transformant comprising the recombinant DNA of (46).
(48) A transformant comprising in its chromosome the DNA of (45).
(49) The transformant according to (47) or (48), which is derived from a coryneform bacterium.
(50) The transformant according to (49), which is derived from Corynebacterium glutamicum.
(51) A method for producing L-lysine, comprising:
culturing the transformant of any one of (47) to (50) in a medium to produce and accumulate L-lysine in the medium, and recovering the L-lysine from the culture.
(52) A method for breeding a coryneform bacterium using the nucleotide sequence information represented by SEQ ID NOS:1 to 3431, comprising the following:
(i) comparing a nucleotide sequence of a genome or gene of a production strain derived a coryneform bacterium which has been subjected to mutation breeding 50 as the produce at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogous thereof by a fermentation method with a corresponding nucleotide sequence in SEQ ID NOS:1 to 3431;
(ii) identifying a mutation point present in the production strain based on a result obtained by the
(iii) introducing the mutation point into a coryneform bacterium which is free of the mutation point; and
(iv) examining productivity by the fermentation method of the compound selected in (i) of the coryneform bacterium obtained in (iii)
(53) The method according to (52), wherein the gene is a gene encoding an enzyme in a biosynthesis pathway or a signal transmission pathway.
(54) The method according to (52), wherein the mutation point is a mutation point relating to a useful mutation which improves or stabilizes the productivity.
(55) A method for breading a coryneform bacterium using the nucleotide sequence information represented by SEQ ID NOS:1 to 3431, comprising:
(i) comparing a nucleotide sequence of a genome or gene of a production strain derived a coryneform bacterium which has been subjected to mutation breeding so as to produce at least one compound selected from an amino acid, a nucleic acid, a vitamin a saccharide, an organic acid, and analogous thereof by a fermentation method, with a corresponding nucleon of sequence in SEQ ID NOS:1 to 3431;
(ii) identifying a mutation point present in the production strain based on the result obtain by (i);
(iii) deleting a mutation point from a coryneform bacterium having the mutation point; and
(iv) examining productivity by the fermentation method of the compound selected in (i) of the coryneform bacterium obtained in (iii).
(56) The method according to (55), wherein the gene is a gene encoding an enzyme in a biosynthesis pathway or a signal transmission pathway.
(57) The method according to (55), wherein the mutation point is a mutation point which decreases or destabilizes the productivity.
(53) A method for breeding a coryneform bacterium using the nucleotide sequence information represented by SEQ ID NOS:2 to 3431, comprising the following:
(i) identifying an isozyme relating to biosynthesis of at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogous thereof, based on the nucleotide sequence information represented by SEQ ID NOS:2 to 3431;
(ii) classifying the isozyme identified in (i) into an isozyme having the same activity;
(iii) mutating all genes encoding the isozyme having the same activity simultaneously; and
(iv) examining productivity by a fermentation method of the compound selected in (i) of the coryneform bacterium which have been transforms with the gene obtained in (iii)
(59) A method for breeding a coryneform bacterium using the nucleotide sequence information represented by SEQ ID NOS:2 to 3431, comprising the following:
(i) arranging a function information of an open reading frame (ORF) represented by SEQ ID NOS:2 to 3431;
(ii) allowing the arranged ORF to correspond to an enzyme on a known biosynthesis or signal transmission pathway;
(iii) explicating an unknown biosynthesis pathway or signal transmission pathway of a coryneform bacterium in combination with information relating known biosynthesis pathway or signal transmission pathway of a coryneform bacterium;
(iv) comparing the pathway explicated in (iii) with a biosynthesis pathway of a target useful product; and
(v) transgenetically varying a coryneform bacterium based on the nucleotide sequence information to either strengthen a pathway which is judged to be important in the biosynthesis of the target useful product in (iv) or weaken a pathway which is judged not to be important in the biosynthesis of the target useful product in (iv).
(60) A coryneform bacterium, bread by the method of any one of (52) to (59).
(61) The coryneform bacterium according to (60), which is a microorganism belonging to the genus Corynebacterium, the genus Brevibacterium, or the genus Microbacterium.
(62) The coryneform bacterium according to (61), wherein the microorganism belonging to the genus Corynebacterium is selected from the group consisting of Corynebacterium glutamicum, Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium callunae, Corynebacterium herculis, Corynebacterium lilium, Corynebacterium melassecola, Corynebacterium thermoaminogenes, and Corynebacterium ammoniagenes.
(63) A method for producing at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid and an analogue thereof, comprising:
culturing a coryneform bacterium of any one of (60) to (62) in a medium to produce and accumulate at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogues thereof;
recovering the compound from the culture.
(64) The method according to (63), wherein the compound is L-lysine.
(65) A method for identifying a protein relating to useful mutation based on proteome analysis, comprising the following:
(i) preparing
a protein derived from a bacterium of a production strain of a coryneform bacterium which has been subjected to mutation breeding by a fermentation process so as to produce at least one compound selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogues thereof, and
a protein derived from a bacterium of a parent strain of the production strain;
(ii) separating the proteins prepared in (i) by two dimensional electrophoresis;
(iii) detecting the separated proteins, and comparing an expression amount of the protein derived from the production strain with that derived from the parent strain;
(iv) treating the protein showing different expression amounts as a result of the comparison with a peptidase to extract peptide fragments;
(v) analyzing amino acid sequences of the peptide fragments obtained in (iv); and
(vi) comparing the amino acid sequences obtained in (v) with the amino acid sequence represented by SEQ ID NOS:3502 to 7001 to identifying the protein having the amino acid sequences.
As used herein, the term “proteome”, which is a coined word by combining “protein” with “genome”, refers to a method for examining of a gene at the polypeptide level.
(66) The method according to (65), wherein the coryneform bacterium is a microorganism belonging to the genus Corynebacterium, the genus Brevibacterium, or the genus Microbacterium.
(67) The method according to (66), wherein the microorganism belonging to the genus Corynebacterium is selected from the group consisting of Corynebacterium glutamicum, Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium callunae, Corynebacterium herculis, Corynebacterium lilium, Corynebacterium melassecola, Corynebacterium thermoaminogenes, and Corynebacterium ammoniagenes.
(68) A biologically pure culture of Corynebacterium glutamicum AHP-3 (FERN BP-7382).
The present invention will be described below in more detail, based on the determination of the full nucleotide sequence of coryneform bacteria.
1. Determination of Full Nucleotide Sequence of Coryneform Bacteria
The term “coryneform bacteria” as used herein means a microorganism belonging to the genus Corynebacterium, the genus Brevibacterium or the genus Microbacterium as defined in Bergeys Manual of Determinative Bacteriology, 8: 599 (1974).
Examples include Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium callunae, Corynebacterium glutamicum, Corynebacterium herculis, Corynebacterium lilium, Corynebacterium melassecola, Corynebacterium thermoaminogenes, Brevibacterium saccharolyticum, Brevibacterium immariophilum, Brevibacterium roseum, Brevibacterium thiogenitalis, Microbacterium ammoniaphilum, and the like.
Specific examples include Corynebacterium acetoacidophilum ATCC 13870, Corynebacterium acetoglutamicum ATCC 15806, Corynebacterium callunae ATCC 15991, Corynebacterium glutamicum ATCC 13032, Corynebacterium glutamicum ATCC 13060, Corynebacterium glutamicum ATCC 13826 (prior genus and species: Brevibacterium flavum, or Corynebacterium lactofermentum) Corynebacterium glutamicum ATCC 14020 (prior genus and species: Brevibacterium divaricatum), Corynebacterium glutamicum ATCC 13869 (prior genus and species: Brevibacterium lactofermentum), Corynebacterium herculis ATCC 13868, Corynebacterium lilium ATCC 15990, Corynebacterium melassecola ATCC 17965, Corynebacterium thermoaminogenes FERM 9244, Brevibacterium saccharolyticum ATCC 4066, Brevibacterium immariophilum ATCC 14068, Brevibacterium roseum ATCC 13825, Brevibacterium thiogenitalis ATCC 19240, Microbacterium ammoniaphilum ATCC 15354, and the like.
(1) Preparation of Genome DNA of Coryneform Bacteria
Coryneform bacteria can be cultured by a conventional method.
Any of a natural medium and a synthetic medium can be used, so long as it is a medium suitable for efficient culturing of the microorganism, and it contains a carbon source, a nitrogen source, an inorganic salt, and the like which can be assimilated by the microorganism.
In Corynebacterium glutamicum, for example, a BY medium (7 g/l meat extract, 10 g/l peptone, 3 g/l sodium chloride, 5 g/l yeast extract, pH 7.2) containing 1% of glycine and the like can be used. The culturing is carried out at 25 to 35° C. overnight.
After the completion of the culture, the cells are recovered from the culture by centrifugation. The resulting cells are washed with a washing solution.
Examples of the washing solution include STE buffer (10.3% sucrose, 25 mmol/l Tris hydrochloride, 25 mmol/l ethylenediaminetetraacetic acid (hereinafter referred to as “EDTA”), pH 8.0), and the like.
Genome DNA can be obtained from the washed cells according to a conventional method for obtaining genome DNA, namely, lysing the cell wall of the cells using a lysozyme and a surfactant (SDS, etc.), eliminating proteins and the like using a phenol solution and a phenol/chloroform solution, and then precipitating the genome DNA with ethanol or the like. Specifically, the following method can be illustrated.
The washed cells are suspended in a washing solution containing 5 to 20 mg/l lysozyme. After shaking, 5 to 20% SDS is added to lyse the cells. In usual, shaking is gently performed at 25 to 40° C. for 30 minutes to 2 hours. After shaking, the suspension is maintained at 60 to 70° C. for 5 to 15 minutes for the lysis.
After the lysis, the suspension is cooled to ordinary temperature, and 5 to 20 ml of Tris-neutralized phenol is added thereto, followed by gently shaking at room temperature for 15 to 45 minutes.
After shaking, centrifugation (15,000×g, 20 minutes, 20° C.) is carried out to fractionate the aqueous layer.
After performing extraction with phenol/chloroform and extraction with chloroform (twice) in the same manner, 3 mol/l sodium acetate solution (pH 5.2) and isopropanol are added to the aqueous layer at 1/10 times volume and 2 times volume, of the aqueous layer, respectively, followed by gently stirring to precipitate the genome DNA.
The genome DNA is dissolved again in a buffer containing 0.01 to 0.04 mg/ml RNase. As an example of the buffer, TE buffer (10 mmol/l Tris hydrochloride, 1 mol/l EDTA, pH 8.0) can be used. After dissolving, the resultant solution is maintained at 25 to 40° C. for 20 to 50 minutes and then extracted successively with phenol, phenol/chloroform and chloroform as in the above case.
After the extraction, isopropanol precipitation is carried out and the resulting DNA precipitate is washed with 70% ethanol, followed by air drying, and then dissolved in TE buffer to obtain a genome DNA solution.
(2) Production of Shotgun Library
A method for produce a genome DNA library using the genome DNA of the coryneform bacteria prepared in the above (1) include a method described in Molecular Cloning, A laboratory Manual, Second Edition (1989) (hereinafter referred to as “Molecular Cloning, 2nd ed.”). In particular, the following method can be exemplified to prepare a genome DNA library appropriately usable in determining the full nucleotide sequence by the shotgun method.
To 0.01 mg of the genome DNA of the coryneform bacteria prepared in the above (1) a buffer, such as TE buffer or the like, is added to give a total volume of 0.4 ml. Then, the genome DNA is digested into fragments of 1 to 10 kb with a sonicator (Yamato Powersonic Model 50). The treatment with the sonicator is performed at an output off 20 continuously for 5 seconds.
The resulting genome DNA fragments are blunt-ended using DNA blunting kit (manufactured by Takara Shuzo) or the like.
The blunt-ended genome fragments are fractionated by agarose gel or polyacrylamide gel electrophoresis and genome fragments of 1 to 2 kb are Cut out from the gel.
To the gel, 0.2 to 0.5 ml of a buffer for eluting DNA, such as MG elution buffer (0.5 mol/l ammonium acetate, 10 mmol/l magnesium acetate, 1 mmol/l EDTA, 0.1% SDS) or the like, is added, followed by shaking at 25 to 40° C. overnight to elute DNA.
The resulting DNA eluate is treated with phenol/chloroform and then precipitated with ethanol to obtain a genome library insert.
This insert is ligated into a suitable vector, such as pUC18 SmaI/BAP (manufactured by Amersham Pharmacia Biotech) or the like, using T4 ligase (manufactured by Takara Shuzo) or the like. The ligation can be carried out by allowing a mixture to stand at 10 to 20° C. for 20 to 50 hours.
The resulting ligation product is precipitated with ethanol and dissolved in 5 to 20 μl of TE buffer.
Escherichia coli is transformed in accordance with a conventional method using 0.5 to 2 μl of the ligation solution. Examples of the transformation method include the electroporation method using ELECTRO MAX DH10B (manufactured by Life Technologies) for Escherichia coli. The electroporation method can be carved out under the conditions as described in the manufacturer's instructions.
The transformed Escherichia coli is spread on a suitable selection medium containing agar, for example, LB plate medium containing 10 to 100 mg/l ampicillin (LB medium (10 g/l bactotrypton, 5 g/l yeast extract, 10 g/l sodium chloride, pH 7.0) containing 1.6% of agar) when pUC18 is used as the cloning vector, and cultured therein.
The transformant can be obtained as colonies formed on the plate medium. In this step, it is possible to select the transformant having the recombinant DNA containing the genome DNA as white colonies by adding X-gal and IPTG (isopropyl-β-thiogalactopyranoside) to the plate medium.
The transformant is allowed to stand for culturing in a 96-well titer plate to which 0.05 ml of the LB medium containing 0.1 mg/ml of ampicillin has been added in each well. The resulting culture can be used in an experiment of (4) described below. Also, the culture solution can be stored at −80° C. by adding 0.05 ml per well of the LB medium containing 20% glycerol to the culture solution, followed by mixing, and the stored culture solution can be used at any time.
(3) Production of Cosmid Library
The genome DNA (0.1 mg) of the coryneform bacteria prepared in the above (1) is partially digested with a restriction enzyme, such as Sau3AI or the like, and then ultracentrifuged (26,000 rpm, 18 hours, 20° C.) under a 10 to 40% sucrose density gradient using a 10% sucrose buffer (1 mol/l NaCl, 20 mmol/l Tris hydrochloride, 5 mmol/l EDTA, 10% sucrose, pH 8.0) and a 40% sucrose buffer (elevating the concentration of the 10% sucrose buffer to 40%).
After the centrifugation, the thus separated solution is fractionated into tubes in 1 ml per each tube. After confirming the DNA fragment size of each fraction by agarose gel electrophoresis, a fraction rich in DNA fragments of about 40 kb is precipitated with ethanol.
The resulting DNA fragment is ligated to a cosmid vector having a cohesive end which can be ligated to the fragment. When the genome DNA is partially digested with Sau3AI, the partially digested product can be ligated to, for example, the BamHI site of superCos1 (manufactured by Stratagene) in accordance with the manufacture's instructions.
The resulting ligation product is packaged using a packaging extract which can be prepared by a method described in Molecular Cloning, 2nd ed. and then used in transforming Escherichia coli. More specifically, the ligation product is packaged using, for example, a commercially available packaging extract, Gigapack III Gold Packaging Extract (manufactured by Stratagene) in accordance with the manufacture's instructions and then introduced into Escherichia coli XL-1-BlueMR (manufactured by Stratagene) or the like.
The thus transformed Escherichia coli is spread on an LB plate medium containing ampicillin, and cultured therein.
The transformant can be obtained as colonies formed on the plate medium.
The transformant is subjected to standing culture in a 96-well titer plate to which 0.05 ml of the LB medium containing 0.1 mg/ml ampicillin has been added.
The resulting culture can be employed in an experiment of (4) described below. Also, the culture solution can be stored at −80° C. by adding 0.05 ml per well of the LB medium containing 20% glycerol to the culture solution, followed by mixing, and the stored culture solution can be used at any time.
(4) Determination of Nucleotide Sequence
(4-1) Preparation of Template
The full nucleotide sequence of genome DNA of coryneform bacteria can be determined basically according to the whole genome shotgun method (Science, 269: 496-512 (1995) ).
The template used in the whole genome shotgun method can be prepared by PCR using the library prepared in the above (2) (DNA Research, 5: 1-9 (1998)).
Specifically, the template can be prepared as follows.
The clone derived from the whole genome shotgun library is inoculated by using a replicator (manufactured by GENETIX) into each well of a 96-well plate to which 0.08 ml per well of the LB medium containing 0.1 mg/ml ampicillin has been added, followed by stationarily culturing at 37° C. overnight.
Next, the culture solution is transported, using a copy plate (manufactured by Tokken), into each well of a 96-well reaction plate (manufactured by PE Biosystems) to which 0.025 ml per well of a PCR reaction solution has been added using TaKaRa Ex Taq (manufactured by Takara Shuzo). Then, PCR is carried out in accordance with the protocol by Makino et al. (DNA Research, 5: 1-9 (1998)) using GeneAmp PCR System 9700 (manufactured by PE Biosystems) to amplify the inserted fragments.
The excessive primers and nucleotides are eliminated using a kit for purifying a PCR product, and the product is used as the template in the sequencing reaction.
It is also possible to determine the nucleotide sequence using a double-stranded DNA plasmid as a template.
The double-stranded DNA plasmid used as the template can be obtained by the following method.
The clone derived from the whole genome shotgun library is inoculated into each well of a 24- or 96-well plate to which 1.5 ml per well of a 2×YT medium (16 g/l bactotrypton, 10 g/l yeast extract, 5 g/l sodium chloride, pH 7.0) containing 0.05 mg/ml ampicillin has been added, followed by culturing under shaking at 37° C. overnight.
The double-stranded DNA plasmid can be prepared from the culture solution using an automatic plasmid preparing machine KURABO PI-50 (manufactured by Kurabo Industries), a multiscreen (manufactured by Millipore) or the like, according to each protocol.
To purify the plasmid, Biomek 2000 manufactured by Beckman Coulter and the like can be used.
The resulting purified double-stranded DNA plasmid is dissolved in water to give a concentration of about 0.1 mg/ml. Then, it can be used as the template in sequencing.
(4-2) Sequencing Reaction
The sequencing reaction can be carried out according to a commercially available sequence kit or the like. A specific method is exemplified below.
To 6 μl of a solution of ABI PRISM BigDye Terminator Cycle Sequencing, Ready Reaction Kit (manufactured by PE Biosystems), 1 to 2 pmol of an M13 regular direction primer (M13-21) or an M13 reverse direction primer (M13REV) (DNA Research, 5: 1-9 (1998)) and 50 to 200 ng of the template prepared in the above (4-1) (the PCR product or plasmid) to give 10 μl of a sequencing reaction solution.
A dye terminator sequencing reaction (35 to 55 cycles) is carried out using this reaction solution and Gene PCR System 9700 (manufactured by PE Biosystems) or the like. The cycle parameter can be determined in accordance with a commercially available kit, for example, the manufacture's instructions attached with ABI PRISM Big Dye Terminator Cycle Sequencing Ready Reaction Kit.
The sample can be purified using a commercially available product, such as Multi Screen HV plate (manufactured by Millipore) or the like, according to the manufacture's instructions.
The thus purified reaction product is precipitated with ethanol, dried and then used for the analysis. The dried reaction product can be stored in the dark at −30° C. and the stored reaction product can be used at any time.
The dried reaction product can be analyzed using a commercially available sequencer and an analyze according to the manufacture's instructions.
Examples of the commercially available sequencer include ABI PRISM 377 DNA Sequencer (manufactured by PE Biosystems) Example of the analyzer include ABI PRISM 3700 DNA Analyzer (manufactured by PE Biosystems).
(5) Assembly
A software, such as phred (The University of Washington) or the like, can be used as base call for use in analyzing the sequence information obtained in the above (4). A software, such as Cross_Match (The University of Washington) or SPS Cross_Match (manufactured by Southwest Parallel Software) or the like, can be used to mass the vector sequence information.
For the assembly, a software, such as phrap (The University of Washington), SPS phrap (manufactured by Southwest Parallel Software) or the like, can be used.
In the above, analysis and output of the results thereof, a computer such as UNIX, PC, Macintosh, and the like can be used.
Contig obtained by the assembly can be analyzed using a graphical editor such as consed (The University of Washington) or the like.
It is also possible to perform a series of the operations from the base call to the assembly in a lump using a script phredPhrap attached to the consed.
As used herein, software will be understood to also be referred to as a comparator.
(6) Determination of Nucleotide Sequence in Gap Part
Each of the cosmids in the cosmid library constructed in the above (3) is prepared in the same manner as in the preparation of the double-stranded DNA plasmid described in the above (4-1). The nucleotide sequence at the end of the insert fragment of the cosmid is determined using a commercially available kit, such as ABI PRISM BigDye Terminator Cycle Sequencing Ready Reaction Kit (manufactured by PE Biosystems) according to the manufacture's instructions.
About 800 cosmid clones are sequenced at both ends of the inserted fragment to detect a nucleotide sequence in the contig derived from the shotgun sequencing obtained in (5) which is coincident with the sequence. Thus, the chain linkage between respective cosmid clones and respective contigs are clarified, and mutual alignment is carried out. Furthermore, the results are compared with known physical maps to map the cosmids and the contigs. In case of Corynebacterium glutamicum ATCC 13032, a physical map of Mol. Gen. Genet., 252: 255-265 (1996) can be used.
The sequence in the region which cannot be covered with the contigs (gap part) can be determined by the following method.
Clones containing sequences positioned at the ends of the contigs are selected. Among these, a clone wherein only one end of the inserted fragment has been determined is selected and the sequence at the opposite end of the inserted fragment is determined.
A shotgun library clone or a cosmid clone derived therefrom containing the sequences at the respective ends of the inserted fragments in the two contigs is identified and the full nucleotide sequence of the inserted fragment of the clone is determined.
According to this method, the nucleotide sequence of the gap part can be determined.
When no shotgun library clone or cosmid clone covering the gap part is available, primers complementary to the end sequences of the two different contigs are prepared and the DNA fragment in the gap part is amplified. Then, sequencing is performed by the primer walking method using the amplified DNA fragment as a template or by the shotgun method in which the sequence of a shotgun clone prepared from the amplified DNA fragment is determined. Thus, the nucleotide sequence of the above-described region can be determined.
In a region showing a low sequence accuracy, primers are synthesized using AUTOFINISH function and NAVIGATING function of consed (The University of Washington), and the sequence is determined by the primer walking method to improve the sequence accuracy.
Examples of the thus determined nucleotide sequence of the full genome include the full nucleotide sequence of genome of Corynebacterium glutamicum ATCC 13032 represented by SEQ ID NO:1.
(7) Determination of Nucleotide Sequence of Microorganism Genome DNA Using the Nucleotide Sequence Represented by SEQ ID NO:1
A nucleotide sequence of a polynucleotide having a homology of 80% or more with. the full nucleotide sequence of Corynebacterium glutamicum ATCC 13032 represented by SEQ ID NO:1 as determined above can also be determined using the nucleotide sequence represented by SEQ ID NO:1, and the polynucleotide having a nucleotide sequence having a homology of 80% or more with the nucleotide sequence represented by SEQ ID NO:1 of the present invention is within the scope of the present invention. The term “polynucleotide having a nucleotide sequence having a homology of 80% or more with the nucleotide sequence represented by SEQ ID NO:1 of the present invention” is a polynucleotide in which a full nucleotide sequence of the chromosome DNA can be determined using as a primer an oligonucleotide composed of continuous 5 to 50 nucleotides in the nucleotide sequence represented by SEQ ID NO:1, for example, according to PCR using the chromosome DNA as a template. A particularly preferred primer in determination of the full nucleotide sequence is an oligonucleotide having nucleotide sequences which are positioned at the interval of about 300 to 500 bp, and among such oligonucleotides, an oligonucleotide having a nucleotide sequence selected from DNAs encoding a protein relating to a main metabolic pathway is particularly preferred. The polynucleotide in which the full nucleotide sequence of the chromosome DNA can be determined using the oligonucleotide includes polynucleotides constituting a chromosome DNA derived from a microorganism belonging to coryneform bacteria. Such a polynucleotide is preferably a polynucleotide constituting chromosome DNA derived from a microorganism belonging to the genus Corynebacterium, more preferably a polynucleotide constituting a chromosome DNA of Corynebacterium glutamicum.
2. Identification of ORF (Open Reading Frame) and Expression Regulatory Fragment and Determination of the Function of ORF
Based on the full nucleotide sequence data of the genome derived from coryneform bacteria determined in the above item 1, an ORF and an expression modulating fragment can be identified. Furthermore, the function of the thus determined ORF can be determined.
The ORF means a continuous region in the nucleotide sequence of mRNA which can be translated as an amino acid sequence to mature to a protein. A region of the DNA coding for the ORF of mRNA is also called ORF.
The expression modulating fragment (hereinafter referred to as “EMF”) is used herein to define a series of polynucleotide fragments which modulate the expression of the ORF or another sequence ligated operatably thereto. The expression “modulate the expression of a sequence ligated operatably” is used herein to refer to changes in the expression of a sequence due to the presence of the EMF. Examples of the EMF include a promoter, an operator, an enhancer, a silencer, a ribosome-binding sequence, a transcriptional termination sequence, and the like. In coryneform bacteria, an EMF is usually present in an intergenic segment (a fragment positioned between two genes; about 10 to 200 nucleotides in length). Accordingly, an EMF is frequently present in an intergenic segment of 10 nucleotides or longer. It is also possible to determine or discover the presence of an EMF by using known EMF sequences as a target sequence or a target structural motif (or a target motif) using an appropriate software or comparator, such as FASTA (Proc. Natl. Acad. Sci. USA, 85: 2444-48 (1988)), BLAST (J. Mol. Biol., 215: 403-410 (1990)) or the like. Also, it can be identified and evaluated using a known EMF-capturing vector (for example, pKK232-8; manufactured by Amersham Pharmacia Biotech).
The term “target sequence” is used herein to refer to a nucleotide sequence composed of 6 or more nucleotides, an amino acid sequence composed of 2 or more amino acids, or a nucleotide sequence encoding this amino acid sequence composed of 2 or more amino acids. A longer target sequence appears at random in a data base at the lower possibility. The target sequence is preferably about 10 to 100 amino acid residues or about 30 to 300 nucleotide residues.
The term “target structural motif” or “target motif” is used herein to refer to a sequence or a combination of sequences selected optionally and reasonably. Such a motif is selected on the basis of the three-dimensional structure formed by the folding of a polypeptide by means known to one of ordinary skill in the art. Various motives are known.
Examples of the target motif of a polypeptide include, but are not limited to, an enzyme activity site, a protein-protein interaction site, a signal sequence, and the like. Examples of the target motif of a nucleic acid include a promoter sequence, a transcriptional regulatory factor binding sequence, a hair pin structure, and the like.
Examples of highly useful EMF include a high-expression promoter, an inducible-expression promoter, and the like. Such an EMF can be obtained by positionally determining the nucleotide sequence of a gene which is known or expected as achieving high expression (for example, ribosomal RNA gene: GenBank Accession No. M16175 or Z46753) or a gene showing a desired induction pattern (for example, isocitrate lyase gene induced by acetic acid: Japanese Published Unexamined Patent Application No. 56782/93) via the alignment with the full genome nucleotide sequence determined in the above item 1, and isolating the genome fragment in the upstream part (usually 200 to 500 nucleotides from the translation initiation site). It is also possible to obtain a highly useful EMF by selecting an EMF showing a high expression efficiency or a desired induction pattern from among promoters captured by the EMF-capturing vector as described above.
The ORF can be identified by extracting characteristics common to individual ORFs, constructing a general model based on these characteristics, and measuring the conformity of the subject sequence with the model. In the identification, a software, such as GeneMark (Nuc. Acids. Res., 22: 4756-67 (1994): manufactured by GenePro)), GeneMark.hmm (manufactured by GenePro), GeneHacker (Protein, Nucleic Acid and Enzyme, 42: 3001-07 (1997)), Glimmer (Nuc. Acids. Res., 26: 544-548 (1998): manufactured by The Institute of Genomic Research), or the like, can be used. In using the software, the default (initial setting) parameters are usually used, though the parameters can be optionally changed.
In the above-described comparisons, a computer, such as UNIX, PC, Macintosh, or the like, can be used.
Examples of the ORF determined by the method of the present invention include ORFs having the nucleotide sequences represented by SEQ ID NOS:2 to 3501 present in the genome of Corynebacterium glutamicum as represented by SEQ ID NO:1. In these ORFs, polypeptides having the amino acid sequences represented by SEQ ID NOS:3502 to 7001 are encoded.
The function of an ORF can be determined by comparing the identified amino acid sequence of the ORF with known homologous sequences using a homology searching software or comparator, such as BLAST, FAST, Smith & Waterman (Meth. Enzym., 164: 765 (1988)) or the like on an amino acid data base, such as Swith-Prot, PIR, GenBank-nr-aa, GenPept constituted by protein-encoding domains derived from GenBank data base, OWL or the like.
Furthermore, by the homology searching, the identity and similarity with the amino acid sequences of known proteins can also be analyzed.
With respect of the term “identity” used herein, where two polypeptides each having 10 amino acids are different in the positions of 3 amino acids, these polypeptides have an identity of 70% with each other. In case wherein one of the different 3 amino acids is analogue (for example, leucine and isoleucine), these polypeptides have a similarity of 80%.
As a specific example, Table 1 shows the registration numbers in known data bases of sequences which are judged as having the highest similarity with the nucleotide sequence of the ORF derived from Corynebacterium glutamicum ATCC 13032, genes of these sequences, functions of these genes, and identities thereof compared with known amino acid translation sequences.
Thus, a great number of novel genes derived from coryneform bacteria can be identified by determining the full nucleotide sequence of the genome derived from coryneform bacterium by the means of the present invention. Moreover, the function of the proteins encoded by these genes can be determined. Since coryneform bacteria are industrially highly useful microorganisms, many of the identified genes are industrially useful.
Moreover, the characteristics of respective microorganisms can be clarified by classifying the functions thus determined. As a result, valuable information in breeding is obtained.
Furthermore, from the ORF information derived from coryneform bacteria, the ORF corresponding to the microorganism is prepared and obtained according to the general method as disclosed in Molecular Cloning, 2nd ed. or the like. Specifically, an oligonucleotide having a nucleotide sequence adjacent to the ORF is synthesized, and the ORF can be isolated and obtained using the oligonucleotide as a primer and a chromosome DNA derived from coryneform bacteria as a template according to the general PCR cloning technique. Thus obtained ORF sequences include polynucleotides comprising the nucleotide sequence represented by any one of SEQ ID NOS:2 to 3501.
The ORF or primer can be prepared using a polypeptide synthesizer based on the above sequence information.
Examples of the polynucleotide of the present invention include a polynucleotide containing the nucleotide sequence of the ORF obtained in the above, and a polynucleotide which hybridizes with the polynucleotide under stringent conditions.
The polynucleotide of the present invention can be a single-stranded DNA, a double-stranded DNA and a single-stranded RNA, though it is not limited thereto.
The polynucleotide which hybridizes with the polynucleotide containing the nucleotide sequence of the ORF obtained in the above under stringent conditions includes a degenerated mutant of the ORF. A degenerated mutant is a polynucleotide fragment having a nucleotide sequence which is different from the sequence of the ORF of the present invention which encodes the same amino acid sequence by degeneracy of a gene code.
Specific examples include a polynucleotide comprising the nucleotide sequence represented by any one of SEQ ID NOS:2 to 3431, and a polynucleotide which hybridizes with the polynucleotide under stringent conditions.
A polynucleotide which hybridizes under stringent conditions is a polynucleotide obtained by colony hybridization, plaque hybridization, Southern blot hybridization or the like using, as a probe, the polynucleotide having the nucleotide sequence of the ORF identified in the above. Specific examples include a polynucleotide which can be identified by carrying out hybridization at 65° C. in the presence of 0.7-1.0 M NaCl using a filter on which a polynucleotide prepared from colonies or plaques is immobilized, and then washing the filter with 0.1× to 2×SSC solution (the composition of 1×SSC contains 150 mM sodium chloride and 15 mM sodium citrate) at 65° C.
The hybridization can be carried out in accordance with known methods described in, for example, Molecular Cloning, 2nd ed., Current Protocols in Molecular Biology, DNA Cloning 1: Core Techniques, A Practical Approach, Second Edition, Oxford University (1995) or the like. Specific examples of the polynucleotide which can be hybridized include a DNA having a homology of 60% or more, preferably 80% or more, and particularly preferably 95% or more, with the nucleotide sequence represented by any one of SEQ ID NO:2 to 3431 when calculated using default (initial setting) parameters of a homology searching software, such as BLAST, FASTA, Smith-Waterman or the like.
Also, the polynucleotide of the present invention includes a polynucleotide encoding a polypeptide comprising the amino acid sequence represented by any one of SEQ ID NOS:3502 to 6931 and a polynucleotide which hybridizes with the polynucleotide under stringent conditions.
Furthermore, the polynucleotide of the present invention includes a polynucleotide which is present in the 5′ upstream or 3′ downstream region of a polynucleotide comprising the nucleotide sequence of any one of SEQ ID NOS:2 to 3431 in a polynucleotide comprising the nucleotide sequence represented by SEQ ID NO:1, and has an activity of regulating an, expression of a polypeptide encoded by the polynucleotide. Specific examples of the polynucleotide having an activity of regulating an expression of a polypeptide encoded by the polynucleotide includes a polynucleotide encoding the above described EMF, such as a promoter, an operator, an enhancer, a silencer, a ribosome-binding sequence, a transcriptional termination sequence, and the like.
The primer used for obtaining the ORF according to the above PCR cloning technique includes an oligonucleotide comprising a sequence which is the same as a sequence of 10 to 200 continuous nucleotides in the nucleotide sequence of the ORF and an adjacent region or an oligonucleotide comprising a sequence which is complementary to the oligonucleotide. Specific examples include an oligonucleotide comprising a sequence which is the same as a sequence of 10 to 200 continuous nucleotides of the nucleotide sequence represented by any one of SEQ ID NOS:1 to 3431, and an oligonucleotide comprising a sequence complementary to the oligonucleotide comprising a sequence of at least 10 to 20 continuous nucleotide of any one of SEQ ID NOS:1 to 3431. When the primers are used as a sense primer and an antisense primer, the above-described oligonucleotides in which melting temperature (Tm) and the number of nucleotides are not significantly different from each other are preferred.
The oligonucleotide of the present invention includes an oligonucleotide comprising a sequence which is the same as 10 to 200 continuous nucleotides of the nucleotide sequence represented by any one of SEQ ID NOS:1 to 3431 or an oligonucleotide comprising a sequence complementary to the oligonucleotide.
Also, analogues of these oligonucleotides (hereinafter also referred to as “analogous oligonucleotides”) are also provided by the present invention and are useful in the methods described herein.
Examples of the analogous oligonucleotides include analogous oligonucleotides in which a phosphodiester bond in an oligonucleotide is converted to a phosphorothioate bond, analogous oligonucleotides in which a phosphodiester bond in an oligonucleotide is converted to an N3′-P5′ phosphoamidate bond, analogous oligonucleotides in which ribose and a phosphodiester bond in an oligonucleotide is converted to a peptide nucleic acid bond, analogous oligonucleotides in which uracil in an oligonucleotide is replaced with C-5 propynyluracil, analogous oligonucleotides in which uracil in an oligonucleotide is replaced with C-5 thiazoluracil, analogous oligonucleotides in which cytosine in an oligonucleotide is replaced with C-5 propynylcytosine, analogous oligonucleotides in which cytosine in an oligonucleotide is replaced with phenoxazine-modified cytosine, analogous oligonucleotides in which ribose in an oligonucleotide is replaced with 2′-O-propylribose, analogous oligonucleotides in which ribose in an oligonucleotide is replaced with 2′-methoxyethoxyribose, and the like (Cell Engineering, 16: 1463 (1997)).
The above oligonucleotides and analogous oligonucleotides of the present invention can be used as probes for hybridization and antisense nucleic acids described below in addition to as primers.
Examples of a primer for the antisense nucleic acid techniques known in the art include an oligonucleotide which hybridizes the oligonucleotide of the present invention under stringent conditions and has an activity regulating expression of the polypeptide encoded by the polynucleotide, in addition to the above oligonucleotide.
3. Determination of Isozymes
Many mutants of coryneform bacteria which are useful in th production of useful substances, such as amino acids, nucleic acids, vitamins, saccharides, organic acids, and the like, are obtained by the present invention.
However, since the gene sequence data of the microorganism has been, to date, insufficient, useful mutants have been obtained by mutagenic techniques using a mutagen, such as nitrosoguanidine (NTG) or the like.
Although genes can be mutated randomly by the mutagenic method using the above-described mutagen, all genes encoding respective isozymes having similar properties relating to the metabolism of intermediates cannot be mutated. In the mutagenic method using a mutagen, genes are mutated randomly. Accordingly, harmful mutations worsening culture characteristics, such as delay in growth, accelerated foaming, and the like, might be imparted at a great frequency, in a random manner.
However, if gene sequence information is available, such as is provided by the present invention, it is possible to mutate all of the genes encoding target isozymes. In this case, harmful mutations may be avoided and the target mutation can be incorporated.
Namely, an accurate number and sequence information of the target isozymes in coryneform bacteria can be obtained based on the ORF data obtained in the above item 2. By using the sequence information, all of the target isozyme genes can be mutated into genes having the desired properties by, for example, the site-specific mutagenesis method described in Molecular Cloning, 2nd ed. to obtain useful mutants having elevated productivity of useful substances.
4. Clarification or Determination of Biosynthesis Pathway and Signal Transmission Pathway
Attempts have been made to elucidate biosynthesis pathways and signal transmission pathways in a number of organisms, and many findings have been reported. However, there are many unknown aspects of coryneform bacteria since a number of genes have not been identified so far.
These unknown points can be clarified by the following method.
The functional information of ORF derived from coryneform bacteria as identified by the method of above item 2 is arranged. The term “arranged” means that the ORF is classified based on the biosynthesis pathway of a substance or the signal transmission pathway to which the ORF belongs using known information according to the functional information. Next, the arranged ORF sequence information is compared with enzymes on the biosynthesis pathways or signal transmission pathways of other known organisms. The resulting information is combined with known data on coryneform bacteria. Thus, the biosynthesis pathways and signal transmission pathways in coryneform bacteria, which have been unknown so far, can be determined.
As a result that these pathways which have been unknown or unclear hitherto are clarified, a useful mutant for producing a target useful substance can be efficiently obtained.
When the thus clarified pathway is judged as important in the synthesis of a useful product, a useful mutant can be obtained by selecting a mutant wherein this pathway has been strengthened. Also, when the thus clarified pathway is judged as not important in the biosynthesis of the target useful product, a useful mutant can be obtained by selecting a mutant wherein the utilization frequency of this pathway is lowered.
5. Clarification or Determination of Useful Mutation Point
Many useful mutants of coryneform bacteria which are suitable for the production of useful substances, such as amino acids, nucleic acids, vitamins, saccharides, organic acids, and the like, have been obtained. However, it is hardly known which mutation point is imparted to a gene to improve the productivity.
However, mutation points contained in production strains can be identified by comparing desired sequences of the genome. DNA of the production strains obtained from coryneform bacteria by the mutagenic technique with the nucleotide sequences of the corresponding genome DNA and ORF derived from coryneform bacteria determined by the methods of the above items 1 and 2 and analyzing them
Moreover, effective mutation points contributing to the production can be easily specified from among these mutation points on the basis of known information relating to the metabolic pathways, the metabolic regulatory mechanisms, the structure activity correlation of enzymes, and the like.
When any efficient mutation can be hardly specified based on known data, the mutation points thus identified can be introduced into a wild strain of coryneform bacteria or a production strain free of the mutation. Then, it is examined whether or not any positive effect can be achieved on the production.
For example, by comparing the nucleotide sequence of homoserine dehydrogenase gene hom of a lysine-producing B-6-strain of Corynebacterium glutamicum (Appl. Microbiol. Biotechnol., 32: 269-273 (1989)) with the nucleotide sequence corresponding to the genome of Corynebacterium glutamicum ATCC 13032 according to the present invention, a mutation of amino acid replacement in which valine at the 59-position is replaced with alanine (Val59Ala) was identified. A strain obtained by introducing this mutation into the ATCC 13032 strain by the gene replacement method can produce lysine, which indicates that this mutation is an effective mutation contributing to the production of lysine.
Similarly, by comparing the nucleotide sequence of pyruvate carboxylase gene pyc of the B-6 strain with the nucleotide sequence corresponding to the ATCC 13032 genome, a mutation of amino acid replacement in which proline at the 458-position was replaced with serine (Pro458Ser) was identified. A strain obtained by introducing this mutation into a lysine-producing strain of No. 58 (FERM BP-7134) of Corynebacterium glutamicum free of this mutation shows an improved lysine productivity in comparison with the No. 58 strain, which indicates that this mutation is an effective mutation contributing to the production of lysine.
In addition, a mutation Ala213Thr in glucose-6-phosphate dehydrogenase was specified as an effective mutation relating to the production of lysine by detecting glucose-6-phosphate dehydrogenase gene zwf of the B-6 strain.
Furthermore, the lysine-productivity of Corynebacterium glutamicum was improved by replacing the base at the 932-position of aspartokinase gene lysC of the Corynebacterium glutamicum ATCC 13032 genome with cytosine to thereby replace threonine at the 311-position by isoleucine, which indicates that this mutation is an effective mutation contributing to the production of lysine.
Also, as another method to examine whether or not the identified mutation point is an effective mutation, there is a method in which the mutation possessed by the lysine-producing strain is returned to the sequence of a wild type strain by the gene replacement method and whether or not it has a negative influence on the lysine productivity. For example, when the amino acid replacement mutation Val59Ala possessed by hom of the lysine-producing B-6 strain was returned to a wild type amino acid sequence, the lysine productivity was lowered in comparison with the B-6 strain. Thus, it was found that this mutation is an effective mutation contributing to the production of lysine.
Effective mutation points can be more efficiently and comprehensively extracted by combining, if needed, the DNA array analysis or proteome analysis described below.
6. Method of Breeding Industrially Advantageous Production Strain
It has been a general practice to construct production strains, which are used industrially in the fermentation production of the target useful substances, such as amino acids, nucleic acids, vitamins, saccharides, organic acids, and the like, by repeating mutagenesis and breeding based on random mutagenesis using mutagens, such as NTG or the like, and screening.
In recent years, many examples of improved production strains have been made through the use of recombinant DNA techniques. In breeding, however, most of the parent production strains to be improved are mutants obtained by a conventional mutagenic procedure (W. Leuchtenberger, Amino Acids—Technical Production and Use. In: Roehr (ed) Biotechnology, second edition, vol. 6, products of primary metabolism. VCH Verlagsgesellschaft mbH, Weinheim, P 465 (1996)).
Although mutagenesis methods have largely contributed to the progress of the fermentation industry, they suffer from a serious problem of multiple, random introduction of mutations into every part of the chromosome. Since many mutations are accumulated in a single chromosome each time a strain is improved, a production strain obtained by the random mutation and selecting is generally inferior in properties (for example, showing poor growth, delayed consumption of saccharides, and poor resistance to stresses such as temperature and oxygen) to a wild type strain, which brings about troubles such as failing to establish a sufficiently elevated productivity, being frequently contaminated with miscellaneous bacteria, requiring troublesome procedures in culture maintenance, and the like, and, in its turn, elevating the production cost in practice. In addition, the improvement in the productivity is based on random mutations and thus the mechanism thereof is unclear. Therefore, it is very difficult to plan a rational breeding strategy for the subsequent improvement in the productivity.
According to the present invention, effective mutation points contributing to the production can be efficiently specified from among many mutation points accumulated in the chromosome of a production strain which has been bred from coryneform bacteria and, therefore, a novel breeding method of assembling these effective mutations in the coryneform bacteria can be established. Thus, a useful production strain can be reconstructed. It is also possible to construct a useful production strain from a wild type strain.
Specifically, a useful mutant can be constructed in the following manner.
One of the mutation points is incorporated into a wild type strain of coryneform bacteria. Then, it is examined whether or not a positive effect is established on the production. When a positive effect is obtained, the mutation point is saved. When no effect is obtained, the mutation point is removed. Subsequently, only a strain having the effective mutation point is used as the parent strain, and the same procedure is repeated. In general, the effectiveness of a mutation positioned upstream cannot be clearly evaluated in some cases when there is a rate-determining point in the downstream of a biosynthesis pathway. It is therefore preferred to successively evaluate mutation points upward from downstream.
By reconstituting effective mutations by the method as described above in a wild type strain or a strain which has a high growth speed or the same ability to consume saccharides as the wild type strain, it is possible to construct an industrially advantageous strain which is free of troubles in the previous methods as described above and to conduct fermentation production using such strains within a short time or at a higher temperature.
For example, a lysine-producing mutant B-6 (Appl. Microbiol. Biotechnol., 32: 262-273 (1989)), which is obtained by multiple rounds of random mutagenesis from a wild type strain Corynebacterium glutamicum ATCC 13032, enables lysine fermentation to be performed at a temperature between 30 and 34° C. but shows lowered growth and lysine productivity at a temperature exceeding 34° C. Therefore, the fermentation temperature should be maintained at 34° C. or lower. In contrast thereto, the production strain described in the above item 5, which is obtained by reconstituting effective mutations relating to lysine production, can achieve a productivity at 40 to 42° C. equal or superior to the result obtained by culturing at 30 to 34° C. Therefore, this strain is industrially advantageous since it can save the load of cooling during the fermentation.
When culture should be carried out at a high temperature exceeding 43° C., a production strain capable of conducting fermentation production at a high temperature exceeding 43° C. can be obtained by reconstituting useful mutations in a, microorganism belonging to the genus Corynebacterium which can grow at high temperature exceeding 43° C. Examples of the microorganism capable of growing at a high temperature exceeding 43° C. include Corynebacterium thermoaminogenes, such as Corynebacterium thermoaminogenes FERM 9244, FERM 9245, FERM 9246 and FERM 9247.
A strain having a further improved productivity of the target product can be obtained using the thus reconstructed strain as the parent strain and further breeding it using the conventional mutagenesis method, the gene amplification method, the gene replacement method using the recombinant DNA technique, the transduction method or the cell fusion method. Accordingly, the microorganism of the present invention includes, but is not limited to, a mutant, a cell fusion strain, a transformant, a transductant or a recombinant strain constructed by using recombinant DNA techniques, so long as it is a producing strain obtained via the step of accumulating at least two effective mutations in a coryneform bacteria in the course of breeding.
When a mutation point judged as being harmful to the growth or production is specified, on the other hand, it is examined whether or not the producing strain used at present contains the mutation point. When it has the mutation, it can be returned to the wild type gene and thus a further useful production strain can be bred.
The breeding method as described above is applicable to microorganisms, other than coryneform bacteria, which have industrially advantageous properties (for example, microorganisms capable of quickly utilizing less expensive carbon sources, microorganisms capable of growing at higher temperatures).
7. Production and Utilization of Polynucleotide Array
(1) Production of Polynucleotide Array
A polynucleotide array can be produced using the polynucleotide or oligonucleotide of the present invention obtained in the above items 1 and 2.
Examples include a polynucleotide array comprising a, solid support to which at least one of a polynucleotide comprising the nucleotide sequence represented by SEQ ID NOS:2 to 3501, a polynucleotide which hybridizes with the polynucleotide under stringent conditions, and a polynucleotide comprising 10 to 200 continuous nucleotides in the nucleotide sequence of the polynucleotide is adhered; and a polynucleotide array comprising a solid support to which at least one of a polynucleotide encoding a polypeptide comprising the amino acid sequence represented by any one of SEQ ID NOS:3502 to 7001, a polynucleotide which hybridizes with the polynucleotide under stringent conditions, and a polynucleotide comprising 10 to 200 continuous bases in the nucleotide sequences of the polynucleotides is adhered.
Polynucleotide arrays of the present invention include substrates known in the art, such as a DNA chip, a DNA microarray and a DNA macroarray, and the like, and comprises a solid support and plural polynucleotides or fragments thereof which are adhered to the surface of the solid support.
Examples of the solid support include a glass plate, a nylon membrane, and the like.
The polynucleotides or fragments thereof adhered to the surface of the solid support can be adhered to the surface of the solid support using the general technique for preparing arrays. Namely, a method in which they are adhered to a chemically surface-treated solid support, for example, to which a polycation such as polylysine or the like has been adhered (Nat. Genet., 21: 15-19 (1999)). The chemically surface-treated supports are commercially available and the commercially available solid product can be used, as the solid support of the polynucleotide array according to the present invention.
As the polynucleotides or oligonucleotides adhered to the solid support, the polynucleotides and oligonucleotides of the present invention obtained in the above items 1 and 2 can be used.
The analysis described below can be efficiently performed by adhering the polynucleotides or oligonucleotides to the solid support at a high density, though a high fixation density is not always necessary.
Apparatus for achieving a high fixation density, such as an arrayer robot or the like, is commercially available from Takara Shuzo (GMS417 Arrayer), and the commercially available product can be used.
Also, the oligonucleotides of the present invention can be synthesized directly on the solid support by the photolithography method or the like (Nat. Genet., 21: 20-24 (1999)). In this method, a linker having a protective group which can be removed by light irradiation is first adhered to a solid support, such as a slide glass or the like. Then, it is irradiated with light through a mask (a photolithograph mask) permeating light exclusively at a definite part of the adhesion part. Next, an oligonucleotide having a protective group which can be removed by light irradiation is added to the part. Thus, a ligation reaction with the nucleotide arises exclusively at the irradiated part. By repeating this procedure, oligonucleotides, each having a desired sequence, different from each other can be synthesized in respective parts. Usually, the oligonucleotides to be synthesized have a length of 10 to 30 nucleotides.
(2) Use of Polynucleotide Array
The following procedures (a) and (b) can be carried out using the polynucleotide array prepared in the above (1).
(a) Identification of Mutation Point of Coryneform Bacterium Mutant and Analysis of Expression Amount and Expression Profile of Gene Encoded by Genome
By subjecting a gene derived from a mutant of coryneform bacteria or an examined gene to the following steps (i) to (iv), the mutation point of the gene can be identified or the expression amount and expression profile of the gene can be analyzed:
(i) producing a polynucleotide array by the method of the above (1);
(ii) incubating polynucleotides immobilized on the polynucleotide array together with the labeled gene derived from a mutant of the coryneform bacterium using the polynucleotide array produced in the above (i) under hybridization conditions;
(iii) detecting the hybridization; and
(iv) analyzing the hybridization data.
The gene derived from a mutant of coryneform bacteria or the examined gene include a gene relating to biosynthesis of at least one selected from amino acids, nucleic acids, vitamins, saccharides, organic acids, and analogues thereof.
The method will be described in detail.
A single nucleotide polymorphism (SNP) in a human region of 2,300 kb has been identified using polynucleotide arrays (Science, 280: 1077-82 (1998)). In accordance with the method of identifying SNP and methods described in Science, 278: 680-686 (1997); Proc. Natl. Acad. Sci. USA, 96: 12833-38 (1999); Science, 284: 1520-23 (1999), and the like using the polynucleotide array produced in the above (1) and a nucleic acid molecule (DNA, RNA) derived from coryneform bacteria in the method of the hybridization, a mutation point of a useful mutant, which is useful in producing an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, or the like can be identified and the gene expression amount and the expression profile thereof can be analyzed.
The nucleic acid molecule (DNA, RNA) derived from the coryneform bacteria can be obtained according to the general method described in Molecular Cloning, 2nd ed. or the like. mRNA derived from Corynebacterium glutamicum can also be obtained by the method of Bormann et al. (Molecular Microbiology, 6: 317-326 (1992)) or the like.
Although ribosomal RNA (rRNA) is usually obtained in large excess in addition to the target mRNA, the analysis is not seriously disturbed thereby.
The resulting nucleic acid molecule derived from coryneform bacteria is labeled. Labeling can be carried out according to a method using a fluorescent dye, a method using a radioisotope or the like.
Specific examples include a labeling method in which psoralen-biotin is crosslinked with RNA extracted from a microorganism and, after hybridization reaction, a fluorescent dye having streptoavidin bound thereto is bound to the biotin moiety (Nat. Biotechnol., 16: 45-48 (1998)); a labeling method in which a reverse transcription reaction is carried out using RNA extracted from a microorganism as a template and random primers as primers, and dUTP having a fluorescent dye (for example, Cy3, Cy5) (manufactured by Amersham Pharmacia Biotech) is incorporated into cDNA (Proc. Natl. Acad. Sci. USA, 96: 12833-38 (1999)); and the like.
The labeling specificity can be improved by replacing the random primers by sequences complementary to the 3′-end of ORF (J. Bacteriol., 181: 6425-40 (1999)).
In the hybridization method, the hybridization and subsequent washing can be carried out by the general method (Nat. Biotechnol., 14: 1675-80 (1996), or the like).
Subsequently, the hybridization intensity is measured depending on the hybridization amount of the nucleic acid molecule used in the labeling. Thus, the mutation point can be identified and the expression amount of the gene can be calculated.
The hybridization intensity can be measured by visualizing the fluorescent signal, radioactivity, luminescence dose, and the like, using a laser confocal microscope, a CCD camera, a radiation imaging device (for example, STORM manufactured by Amersham Pharmacia Biotech), and the like, and then quantifying the thus visualized data.
A polynucleotide array on a solid support can also be analyzed and quantified using a commercially available apparatus, such as GMS418 Array Scanner (manufactured by Takara Shuzo) or the like.
The gene expression amount can be analyzed using a commercially available software (for example, ImaGene manufactured by Takara Shuzo; Array Gauge manufactured by Fuji Photo Film; ImageQuant manufactured by Amersham Pharmacia Biotech, or the like).
A fluctuation in the expression amount of a specific gene can be monitored using a nucleic acid molecule obtained in the time course of culture as the nucleic acid molecule derived from coryneform bacteria. The culture conditions can be optimized by analyzing the fluctuation.
The expression profile of the microorganism at the total gene level (namely, which genes among a great number of genes encoded by the genome have been expressed and the expression ratio thereof) can be determined using a nucleic acid molecule having the sequences of many genes determined from the full genome sequence of the microorganism. Thus, the expression amount of the genes determined by the full genome sequence can be analyzed and, in its turn, the biological conditions of the microorganism can be recognized as the expression pattern at the full gene level.
(b) Confirmation of the Presence of Gene Homologous to Examined Gene in Coryneform Bacteria
Whether or not a gene homologous to the examined gene, which is present in an organism other than coryneform bacteria, is present in coryneform bacteria can be detected using the polynucleotide array prepared in the above (1).
This detection can be carried out by a method in which an examined gene which is present in an organism other than coryneform bacteria is used instead of the nucleic acid molecule derived from coryneform bacteria used in the above identification/analysis method of (1).
8. Recording Medium Storing Full Genome Nucleotide Sequence and ORF Data and Being Readable by a Computer and Methods for Using the Same
The term “recording medium or storage device which is readable by a computer” means a recording medium or storage medium which can be directly readout and accessed with a computer. Examples include magnetic recording media, such as a floppy disk, a hard disk, a magnetic tape, and the like; optical recording media, such as CD-ROM, CD-R, CD-RW, DVD-ROM, DVD-RAM, DVD-RW, and the like; electric recording media, such as RAM, ROM, and the like; and hybrids in these categories (for example, magnetic/optical recording media, such as MO and the like).
Instruments for recording or inputting in or on the recording medium or instruments or devices for reading out the information in the recording medium can be appropriately selected, depending on the type of the recording medium and the access device utilized. Also, various data processing programs, software, comparator and formats are used for recording and utilizing the polynucleotide sequence information or the like of the present invention in the recording medium. The information can be expressed in the form of a binary file, a text file or an ASCII file formatted with commercially available software, for example. Moreover, software for accessing the sequence information is available and known to one of ordinary skill in the art.
Examples of the information to be recorded in the above-described medium include the full genome nucleotide sequence information of coryneform bacteria as obtained in the above item 2, the nucleotide sequence information of ORF, the amino acid sequence information encoded by the ORF, and the functional information of polynucleotides coding for the amino acid sequences.
The recording medium or storage device which is readable by a computer according to the present invention refers to a medium in which the information of the present invention has been recorded. Examples include recording media or storage devices which are readable by a computer storing the nucleotide sequence information represented by SEQ ID NOS:1 to 3501, the amino acid sequence information represented by SEQ ID NOS:3502 to 7001, the functional information of the nucleotide sequences represented by SEQ ID NOS:1 to 3501, the functional information of the amino acid sequences represented by SEQ ID NOS:3502 to 7001, and the information listed in Table 1 below and the like.
9. System Based on a Computer Using the Recording Medium of the Present Invention which is Readable by a Computer
The term “system based on a computer” as used herein refers a system composed of hardware device(s) software device(s), and data recording device(s) which are used for analyzing the data recorded in the recording medium of the present invention which is readable by a computer.
The hardware device(s) are, for example, composed of an input unit, a data recording unit, a central processing unit and an output unit collectively or individually.
By the software device(s), the data recorded in the recording medium of the present invention are searched or analyzed using the recorded data and the hardware device(s) as described herein. Specifically, the software device(s) contain at least one program which acts on or with the system in order to screen, analyze or compare biologically meaningful structures or information from the nucleotide sequences, amino acid sequences and the like recorded in the recording medium according to the present invention.
Examples of the software device(s) for identifying ORF and EMF domains include GeneMark (Nuc. Acids. Res., 22: 4756-67 (1994)), GeneHacker (Protein, Nucleic Acid and Enzyme, 42: 3001-07 (1997)), Glimmer (The Institute of Genomic Research; Nuc. Acids. Res., 26: 544-548 (1998)) and the like. In the process of using such a software device, the default (initial setting) parameters are usually used, although the parameters can be changed, if necessary, in a manner known to one of ordinary skill in the art.
Examples of the software device(s) for identifying a genome domain or a polypeptide domain analogous to the target sequence or the target structural motif (homology searching) include FASTA, BLAST, Smith-Waterman, GenetyxMac (manufactured by Software Development), GCG Package (manufactured by Genetic Computer Group), GenCore (manufactured by Compugen), and the like. In the process of using such a software device, the default (initial setting) parameters are usually used, although the parameters can be changed, if necessary, in a manner known to one of ordinary skill in the art.
Such a recording medium storing the full genome sequence data is useful in preparing a polynucleotide array by which the expression amount of a gene encoded by the genome DNA of coryneform bacteria and the expression profile at the total gene level of the microorganism, namely, which genes among many genes encoded by the genome have been expressed and the expression ratio thereof, can be determined.
The data recording device(s) provided by the present invention are, for example, memory device(s) for recording the data recorded in the recording medium of the present invention and target sequence or target structural motif data, or the like, and a memory accessing device(s) for accessing the same.
Namely, the system based on a computer according to the present invention comprises the following:
(i) a user input device that inputs the information stored in the recording medium of the present invention, and target sequence or target structure motif information;
(ii) a data storage device for at least temporarily storing the input information;
(iii) a comparator that compares the information stored in the recording medium of the present invention with the target sequence or target structure motif information, recorded by the data storing device of (ii) for screening and analyzing nucleotide sequence information which is coincident with or analogous to the target sequence or target structure motif information; and
(iv) an output device that shows a screening or analyzing result obtained by the comparator.
This system is usable in the methods in items 2 to 5 as described above for searching and analyzing the ORF and EMF domains, target sequence, target structural motif, etc. of a coryneform bacterium, searching homologs, searching and analyzing isozymes, determining the biosynthesis pathway and the signal transmission pathway, and identifying spots which have been found in the proteome analysis. The term “homologs” as used herein includes both of orthologs and paralogs.
10. Production of Polypeptide Using ORF Derived from Coryneform Bacteria
The polypeptide of the present invention can be produced using a polynucleotide comprising the ORF obtained in the above item 2. Specifically, the polypeptide of the present invention can be produced by expressing the polynucleotide of the present invention or a fragment thereof in a host cell, using the method described in Molecular Cloning, 2nd ed., Current Protocols in Molecular Biology, and the like, for example, according to the following method.
A DNA fragment having a suitable length containing a part encoding the polypeptide is prepared from the full length ORF sequence, if necessary.
Also, DNA in which nucleotides in a nucleotide sequence at a part encoding the polypeptide of the present invention are replaced to give a codon suitable for expression of the host cell, if necessary. The DNA is useful for efficiently producing the polypeptide of the present invention.
A recombinant vector is prepared by inserting the DNA fragment into the downstream of a promoter in a suitable expression vector.
The recombinant vector is introduced to a host cell suitable for the expression vector.
Any of bacteria, yeasts, animal cells, insect cells, plant cells, and the like can be used as the host cell so long as it can be expressed in the gene of interest.
Examples of the expression vector include those which can replicate autonomously in the above-described host cell or can be integrated into chromosome and have a promoter at such a position that the DNA encoding the polypeptide of the present invention can be transcribed.
When a procaryote cell, such as a bacterium or the like, is used as the host cell, it is preferred that the recombinant vector containing the DNA encoding the polypeptide of the present invention can replicate autonomously in the bacterium and is a recombinant vector constituted by, at least a promoter, a ribosome binding sequence, the DNA of the present invention and a transcription termination sequence. A promoter controlling gene can also be contained therewith in operable combination.
Examples of the expression vectors include a vector plasmid which is replicable in Corynebacterium glutamicum, such as pCG1 (Japanese Published Unexamined Patent Application No. 134500/82), pCG2 (Japanese Published Unexamined Patent Application No. 35197/83), pCG4 (Japanese Published Unexamined Patent Application No. 183799/82), pCG11 (Japanese Published Unexamined Patent Application No. 134500/82), pCG116, pCE54 and pCB101 (Japanese Published Unexamined Patent Application No. 105999/83), pCE51, pCE52 and pCE53 (Mol. Gen. Genet., 196: 175-178 (1984)), and the like; a vector plasmid which is replicable in Escherichia coli, such as pET3 and pET11 (manufactured by Stratagene), pBAD, pThioHis and pTrcHis (manufactured by Invitrogen), pKK223-3 and pGEX2T (manufactured by Amersham Pharmacia Biotech), and the like; and pBTrp2, pBTac1 and pBTac2 (manufactured by Boehringer Mannheim Co.), pSE280 (manufactured by Invitrogen), pGEMEX-1 (manufactured by Promega), pQE-8 (manufactured by QIAGEN), pKYP10 (Japanese Published Unexamined Patent Application No. 110600/83), pKYP200 (Agric. Biol. Chem., 48: 669 (1984)), pLSA1 (Agric. Biol. Chem., 53: 277 (1989)), pGEL1 (Proc. Natl. Acad. Sci. USA, 82: 4306 (1985)), pBluescript II SK(-) (manufactured by Stratagene), pTrs30 (prepared from Escherichia coli JM109/pTrS30 (FERM BP-5407)), pTrs32 (prepared from Escherichia coli JM109/pTrS32 (FERM BP-5408)), pGHA2 (prepared from Escherichia coli IGHA2 (FERM B-400), Japanese Published Unexamined Patent Application No. 221091/85), pGKA2 (prepared from Escherichia coli IGKA2 (FERM BP-6798), Japanese Published Unexamined Patent Application No. 221091/85), pTerm2 (U.S. Pat. Nos. 4,686,191, 4,939,094 and 5,160,735), pSupex, pUB110, pTP5, pC194 and pEG400 (J. Bacteriol., 172: 2392 (1990)), pGEX (manufactured by Pharmacia), pET system (manufactured by Novagen), and the like.
Any promoter can be used so long as it can function in the host cell. Examples include promoters derived from Escherichia coli, phage and the like, such as trp promoter (Ptrp), lac promoter, PL promoter, PR promoter, T7 promoter and the like. Also, artificially designed and modified promoters, such as a promoter in which two Ptrp are linked in series (Ptrp×2), tac promoter, lacT7 promoter letI promoter and the like, can be used.
It is preferred to use a plasmid in which the space between Shine-Dalgarno sequence which is the ribosome binding sequence and the initiation codon is adjusted to an appropriate distance (for example, 6 to 18 nucleotides).
The transcription termination sequence is not always necessary for the expression of the DNA of the present invention. However, it is preferred to arrange the transcription terminating sequence at just downstream of the structural gene.
One of ordinary skill in the art will appreciate that the codons of the above-described elements may be optimized, in a known manner, depending on the host cells and environmental conditions utilized.
Examples of the host cell include microorganisms belonging to the genus Escherichia, the genus Serratia, the genus Bacillus, the genus Brevibacterium, the genus Corynebacterium, the genus Microbacterium, the genus Pseudomonas, and the like. Specific examples include Escherichia coli XL1-Blue, Escherichia coli XL2-Blue, Escherichia coli DH1, Escherichia coli MC1000, Escherichia coli KY3276, Escherichia coli W1485; Escherichia coli JM109, Escherichia coli HB101, Escherichia coli No. 49, Escherichia coli W3110, Escherichia coli NY49, Escherichia coli G1698, Escherichia coli TB1, Serratia ficaria, Serratia fonticola, Serratia liquefaciens, Serratia marcescens, Bacillus subtilis, Bacillus amyloliquefaciens, Corynebacterium ammoniagenes, Brevibacterium immariophilum ATCC 14068, Brevibacterium saccharolyticum ATCC 14066, Corynebacterium glutamicum ATCC 13032, Corynebacterium glutamicum ATCC 13869, Corynebacterium glutamicum ATCC 14067 (prior genus and species: Brevibacterium flavum), Corynebacterium glutamicum ATCC 13869 (prior genus and species: Brevibacterium lactofermentum, or Corynebacterium lactofermentum), Corynebacterium acetoacidophilum ATCC 13870, Corynebacterium thermoaminogenes FERM 9244, Microbacterium ammoniaphilum ATCC 15354, Pseudomonas putida, Pseudomonas sp. D-0110, and the like.
When Corynebacterium glutamicum or an analogous microorganism is used as a host, an EMF necessary for expressing the polypeptide is not always contained in the vector so long as the polynucleotide of the present invention contains an EMF. When the EMF is not contained in the polynucleotide, it is necessary to prepare the EMF separately and ligate it so as to be in operable combination. Also, when a higher expression amount or specific expression regulation is necessary, it is necessary to ligate the EMF corresponding thereto so as to put the EMF in operable combination with the polynucleotide. Examples of using an externally ligated EMF are disclosed in Microbiology, 142: 1297-1309 (1996).
With regard to the method for the introduction of the recombinant vector, any method for introducing DNA into the above-described host cells, such as a method in which a calcium ion is used (Proc. Natl. Acad. Sci. USA, 69: 2110 (1972)), a protoplast method (Japanese Published Unexamined Patent Application No. 2483942/88), the methods described in Gene, 17: 107 (1982) and Molecular & General Genetics, 168: 111 (1979) and the like, can be used.
When yeast is used as the host cell, examples of the expression vector include pYES2 (manufactured by Invitrogen), YEp13 (ATCC 37115), YEp24 (ATCC 37051), YCp50 (ATCC 37419), pHS19, pHS15, and the like.
Any promoter can be used so long as it can be expressed in yeast. Examples include a promoter of a gene in the glycolytic pathway, such as hexose kinase and the like, PHO5 promoter, PGK promoter, GAP promoter, ADH promoter, gal 1 promoter, gal 10 promoter, a heat shock protein promoter, MF α1 promoter, CUP 1 promoter, and the like.
Examples of the host cell include microorganisms belonging to the genus Saccharomyces, the genus Schizosaccharomyces, the genus Kluyveromyces, the genus Trichosporon, the genus Schwanniomyces, the genus Pichia, the genus Candida and the like. Specific examples include Saccharomyces cerevisiae, Schizosaccharomyces pombe, Kluyveromyces lactis, Trichosporon pullulans, Schwanniomyces alluvius, Candida utilis and the like.
With regard to the method for the introduction of the recombinant vector, any method for introducing DNA into yeast, such as an electroporation method (Methods. Enzymol., 194: 182 (1990)), a spheroplast method (Proc. Natl. Acad. Sci. USA, 75: 1929 (1978)), a lithium acetate method (J. Bacteriol., 153: 163 (1983)), a method described in Proc. Natl. Acad. Sci. USA, 75: 1929 (1978) and the like, can be used.
When animal cells are used as the host cells, examples of the expression vector include pcDNA3.1, pSinRep5 and pCEP4 (manufactured by Invitorogen), pRev-Tre (manufactured by Clontech), pAxCAwt (manufactured by Takara Shuzo), pcDNAI and pcDM8 (manufactured by Funakoshi), pAGE107 (Japanese Published Unexamined Patent Application No. 22979/91; Cytotechnology, 3:133 (1990)), pAS3-3 (Japanese Published Unexamined Patent Application No. 227075/90), pcDM8 (Nature, 329: 840 (1987)), pcDNAI/Amp (manufactured by Invitrogen), pREP4 (manufactured by Invitrogen), pAGE103 (J. Biochem., 101: 1307 (1987)), pAGE210, and the like.
Any promoter can be used so long as it can function in animal cells. Examples include a promoter of IE (immediate early) gene of cytomegalovirus (CMV), an early promoter of SV40, a promoter of retrovirus, a metallothionein promoter, a heat shock promoter, SRα promoter, and the like. Also, the enhancer of the IE gene of human CMV can be used together with the promoter.
Examples of the host cell include human Namalwa cell, monkey COS cell, Chinese hamster CHO cell, HST5637 (Japanese Published Unexamined Patent Application No. 299/88), and the like.
The method for introduction of the recombinant vector into animal cells is not particularly limited, so long as it is the general method for introducing DNA into animal cells, such as an electroporation method (Cytotechnology, 3: 133 (1990)), a calcium phosphate method (Japanese Published Unexamined Patent Application No. 227075/90), a lipofection method (Proc. Natl. Acad. Sci. USA, 84, 7413 (1987)), the method described in Virology, 52: 456 (1973), and the like.
When insect cells are used as the host cells, the polypeptide can be expressed, for example, by the method described in Bacurovirus Expression Vectors, A Laboratory Manual, W.H. Freeman and Company, New York (1992), Bio/Technology, 6: 47 (1988), or the like.
Specifically, a recombinant gene transfer vector and bacurovirus are simultaneously inserted into insect cells to obtain a recombinant virus in an insect cell culture supernatant, and then the insect cells are infected with the resulting recombinant virus to express the polypeptide.
Examples of the gene introducing vector used in the method include pBlueBac4.5, pVL1392, pVL1393 and pBlueBacIII (manufactured by Invitrogen), and the like.
Examples of the bacurovirus include Autographa californica nuclear polyhedrosis virus with which insects of the family Barathra are infected, and the like.
Examples of the insect cells include Spodoptera frugiperda oocytes Sf9 and Sf21 (Bacurovirus Expression Vectors, A Laboratory Manual, W.H. Freeman and Company, New York (1992)), Trichoplusia ni oocyte High 5 (manufactured by Invitrogen) and the like.
The method for simultaneously incorporating the above-described recombinant gene transfer vector and the above-described bacurovirus for the preparation of the recombinant virus include calcium phosphate method (Japanese Published Unexamined Patent Application No. 227075/90), lipofection method (Proc. Natl. Acad. Sci. USA, 84: 7413 (1987)) and the like.
When plant cells are used as the host cells, examples of expression vector include a Ti plasmid, a tobacco mosaic virus vector, and the like.
Any promoter can be used so long as it can be expressed in plant cells. Examples include 35S promoter of cauliflower mosaic virus (CaMV), rice actin 1 promoter, and the like.
Examples of the host cells include plant cells and the like, such as tobacco, potato, tomato, carrot, soybean, rape, alfalfa, rice, wheat, barley, and the like.
The method for introducing the recombinant vector is not particularly limited, so long as it is the general method for introducing DNA into plant cells, such as the Agrobacterium method (Japanese Published Unexamined Patent Application No. 140885/84, Japanese Published Unexamined Patent Application No. 70080/85, WO 94/00977), the electroporation method (Japanese Published Unexamined Patent Application, No. 251887/85), the particle gun method (Japanese Patents 2606856 and 2517813), and the like.
The transformant of the present invention includes a transformant containing the polypeptide of the present invention per se rather than as a recombinant vector, that is, a transformant containing the polypeptide of the present invention which is integrated into a chromosome of the host, in addition to the transformant containing the above recombinant vector.
When expressed in yeasts, animal cells, insect cells or plant cells, a glycopolypeptide or glycosylated polypeptide can be obtained.
The polypeptide can be produced by culturing the thus obtained transformant of the present invention in a culture medium to produce and accumulate the polypeptide of the present invention or any polypeptide expressed under the control of an EMF of the present invention, and recovering the polypeptide from the culture.
Culturing of the transformant of the present invention in a culture medium is carried out according to the conventional method as used in culturing of the host.
When the transformant of the present invention is obtained using a prokaryote, such as Escherichia coli or the like, or a eukaryote, such as yeast or the like, as the host, the transformant is cultured.
Any of a natural medium and a synthetic medium can be used, so long as it contains a carbon source, a nitrogen source, an inorganic salt and the like which can be assimilated by the transformant and can perform culturing of the transformant efficiently.
Examples of the carbon source include those which can be assimilated by the transformant, such as carbohydrates (for example, glucose, fructose, sucrose, molasses containing them, starch, starch hydrolysate, and the like), organic acids (for example, acetic acid, propionic acid, and the like), and alcohols (for example, ethanol, propanol, and the like).
Examples of the nitrogen source include ammonia, various ammonium salts of inorganic acids or organic acids (for example, ammonium chloride, ammonium sulfate, ammonium acetate, ammonium phosphate, and the like), other nitrogen-containing compounds, peptone, meat extract, yeast extract, corn steep liquor, casein hydrolysate, soybean meal and soybean meal hydrolysate, various fermented cells and hydrolysates thereof, and the like.
Examples of inorganic salt include potassium dihydrogen phosphate, dipotassium hydrogen phosphate, magnesium phosphate, magnesium sulfate, sodium chloride, ferrous sulfate, manganese sulfate, copper sulfate, calcium carbonate, and the like.
The culturing is carried out under aerobic conditions by shaking culture, submerged-aeration stirring culture or the like. The culturing temperature is preferably from 15 to 40° C., and the culturing time is generally from 16 hours to 7 days. The pH of the medium is preferably maintained at 3.0 to 9.0 during the culturing. The pH can be adjusted using an inorganic or organic acid, an alkali solution, urea, calcium carbonate, ammonia, or the like.
Also, antibiotics, such as ampicillin, tetracycline, and the like, can be added to the medium during the culturing, if necessary.
When a microorganism transformed with a recombinant vector containing an inducible promoter is cultured, an inducer can be added to the medium, if necessary.
For example, isopropyl-β-D-thiogalactopyranoside (IPTG) or the like can be added to the medium when a microorganism transformed with a recombinant vector containing lac promoter is cultured, or indoleacrylic acid (IAA) or the like can by added thereto when a microorganism transformed with an expression vector containing trp promoter is cultured.
Examples of the medium used in culturing a transformant obtained using animal cells as the host cells include RPMI 1640 medium (The Journal of the American Medical Association, 199: 519 (1967)), Eagle's MEM medium (Science, 122: 501 (1952)), Dulbecco's modified MEM medium (Virology, 8, 396 (1959)), 199 Medium (Proceeding of the Society for the Biological Medicine, 73:1 (1950)), the above-described media to which fetal calf serum has been added, and the like.
The culturing is carried out generally at a pH of 6 to 8 and a temperature of 30 to 40° C. in the presence of 5% CO2 for 1 to 7 days.
Also, if necessary, antibiotics, such as kanamycin, penicillin, and the like, can be added to the medium during the culturing.
Examples of the medium used in culturing a transformant obtained using insect cells as the host cells include TNM-FH medium (manufactured by Pharmingen), Sf-900 II SFM (manufactured by Life Technologies), ExCell 400 and ExCell 405 (manufactured by JRH Biosciences), Grace's Insect Medium (Nature, 195: 788 (1962)), and the like.
The culturing is carried out generally at a pH of 6 to 7 and a temperature of 25 to 30° C. for 1 to 5 days.
Additionally, antibiotics, such as gentamicin and the like, can be added to the medium during the culturing, if necessary.
A transformant obtained by using a plant cell as the host cell can be used as the cell or after differentiating to a plant cell or organ. Examples of the medium used in the culturing of the transformant include Murashige and Skoog (MS) medium, White medium, media to which a plant hormone, such as auxin, cytokinine, or the like has been added, and the like.
The culturing is carried out generally at a pH of 5 to 9 and a temperature of 20 to 40° C. for 3 to 60 days.
Also, antibiotics, such as kanamycin, hygromycin and the like, can be added to the medium during the culturing, if necessary.
As described above, the polypeptide can be produced by culturing a transformant derived from a microorganism, animal cell or plant cell containing a recombinant vector to which a DNA encoding the polypeptide of the present invention has been inserted according to the general culturing method to produce and accumulate the polypeptide, and recovering the polypeptide from the culture.
The process of gene expression may include secretion of the encoded protein production or fusion protein expression and the like in accordance with the methods described in Molecular Cloning, 2nd ed., in addition to direct expression.
The method for producing the polypeptide of the present invention includes a method of intracellular expression in a host cell, a method of extracellular secretion from a host cell, or a method of production on a host cell membrane outer envelope. The method can be selected by changing the host cell employed or the structure of the polypeptide produced.
When the polypeptide of the present invention is produced in a host cell or on a host cell membrane outer envelope, the polypeptide can be positively secreted extracellularly according to, for example, the method of Paulson et al. (J. Biol. Chem., 264: 17619 (1989)), the method of Lowe et al. (Proc. Natl. Acad. Sci. USA, 86: 8227 (1989); Genes Develop., 4: 1288 (1990)), and/or the methods described in Japanese Published Unexamined Patent Application No. 336963/93? WO 94/23021, and the like.
Specifically, the polypeptide of the present invention can be positively secreted extracellularly by expressing it in the form that a signal peptide has been added to the foreground of a polypeptide containing an active site of the polypeptide of the present invention according to the recombinant DNA technique.
Furthermore, the amount produced can be increased using a gene amplification system, such as by use of a dihydrofolate reductase gene or the like according to the method described in Japanese Published Unexamined Patent Application No. 227075/90.
Moreover, the polypeptide of the present invention can be produced by a transgenic animal individual (transgenic nonhuman animal) or plant individual (transgenic plant).
When the transformant is the animal individual or plant individual, the polypeptide of the present invention can be produced by breeding or cultivating it so as to produce and accumulate the polypeptide, and recovering the polypeptide from the animal individual or plant individual.
Examples of the method for producing the polypeptide of the present invention using the animal individual include a method for producing the polypeptide of the present invention in an animal developed by inserting a gene according to methods known to those of ordinary skill in the art (American Journal of Clinical Nutrition, 63: 639S (1996), American Journal of Clinical Nutrition, 63: 627S (1996), Bio/Technology, 9: 830 (1991)).
In the animal individual, the polypeptide can be produced by breeding a transgenic nonhuman animal to which the DNA encoding the polypeptide of the present invention has been inserted to produce and accumulate the polypeptide in the animal, and recovering the polypeptide from the animal. Examples of the production and accumulation place in the animal include milk (Japanese Published Unexamined Patent Application No. 309192/88), egg and the like of the animal. Any promoter can be used, so long as it can be expressed in the animal. Suitable examples include an α-casein promoter, a β-casein promoter, a β-lactoglobulin promoter, a whey acidic protein promoter, and the like, which are specific for mammary glandular cells.
Examples of the method for producing the polypeptide of the present invention using the plant individual include a method for producing the polypeptide of the present invention by cultivating a transgenic plant to which the DNA encoding the protein of the present invention by a known method (Tissue Culture, 20 (1994), Tissue Culture, 21 (1994), Trends in Biotechnology, 15: 45 (1997)) to produce and accumulate the polypeptide in the plant, and recovering the polypeptide from the plant.
The polypeptide according to the present invention can also be obtained by translation in vitro.
The polypeptide of the present invention can be produced by a translation system in vitro. There are, for example, two in vitro translation methods which may be used, namely, a method using RNA as a template and another method using DNA as a template. The template RNA includes the whole RNA, mRNA, an in vitro transcription product, and the like. The template DNA includes a plasmid containing a transcriptional promoter and a target gene integrated therein and downstream of the initiation site, a PCR/RT-PCR product and the like. To select the most suitable system for the in vitro translation, the origin of the gene encoding the protein to be synthesized (prokaryotic cell/eucaryotic cell), the type of the template (DNA/RNA), the purpose of using the synthesized protein and the like should be considered. In vitro translation kits having various characteristics are commercially available from many companies (Boehringer Mannheim, Promega, Stratagene, or the like), and every kit can be used in producing the polypeptide according to the present invention.
Transcription/translation of a DNA nucleotide sequence cloned into a plasmid containing a T7 promoter can be carried out using an in vitro transcription/translation system E. coli T7 S30 Extract System for Circular DNA (manufactured by Promega, catalogue No. L1130). Also, transcription/translation using, as a template, a linear prokaryotic DNA of a supercoil non-sensitive promoter, such as lacUV5, tac, λPL(con), λPL, or the like, can be carried out using an in vitro transcription/translation system E. coli S30 Extract System for Linear Templates (manufactured by Promega, catalogue No. L1030). Examples of the linear prokaryotic DNA used as a template include a DNA fragment, a PCR-amplified DNA product, a duplicated oligonucleotide ligation, an in vitro transcriptional RNA, a prokaryotic RNA, and the like.
In addition to the production of the polypeptide according to the present invention, synthesis of a radioactive labeled protein, confirmation of the expression capability of a cloned gene, analysis of the function of transcriptional reaction or translation reaction, and the like can be carried out using this system.
The polypeptide produced by the transformant of the present invention can be isolated and purified using the general method for isolating and purifying an enzyme. For example, when the polypeptide of the present invention is expressed as a soluble product in the host cells, the cells are collected by centrifugation after cultivation, suspended in an aqueous buffer, and disrupted using an ultrasonicator, a French press, a Manton Gaulin homogenizer, a Dynomill, or the like to obtain a cell-free extract. From the supernatant obtained by centrifuging the cell-free extract, a purified product can be obtained by the general method used for isolating and purifying an enzyme, for example, solvent extraction, salting out using ammonium sulfate or the like, desalting, precipitation using an organic solvent, anion exchange chromatography using a resin, such as diethylaminoethyl (DEAE)-Sepharose, DIAION HPA-75 (manufactured by Mitsubishi Chemical) or the like, cation exchange chromatography using a resin, such as S-Sepharose FF (manufactured by Pharmacia) or the like, hydrophobic chromatography using a resin, such as butyl sepharose, phenyl sepharose or the like, gel filtration using a molecular sieve, affinity chromatography, chromatofocusing, or electrophoresis, such as isoelectronic focusing or the like, alone or in combination thereof.
When the polypeptide is expressed as an insoluble product in the host cells, the cells are collected in the same manner, disrupted and centrifuged to recover the insoluble product of the polypeptide as the precipitate fraction. Next, the insoluble product of the polypeptide is solubilized with a protein denaturing agent. The solubilized solution is diluted or dialyzed to lower the concentration of the protein denaturing agent in the solution. Thus, the normal configuration of the polypeptide is reconstituted. After the procedure, a purified product of the polypeptide can be obtained by a purification/isolation method similar to the above.
When the polypeptide of the present invention or its derivative (for example, a polypeptide formed by adding a sugar chain thereto) is secreted out of cells, the polypeptide or its derivative can be collected in the culture supernatant. Namely, the culture supernatant is obtained by treating the culture medium in a treatment similar to the above (for example, centrifugation). Then, a purified product can be obtained from the culture medium using a purification/isolation method similar to the above.
The polypeptide obtained by the above method is within the scope of the polypeptide of the present invention, and examples include a polypeptide encoded by a polynucleotide comprising the nucleotide sequence selected from SEQ ID NOS:2 to 3431, and a polypeptide comprising an amino acid sequence represented by any one of SEQ ID NOS:3502 to 6931.
Furthermore, a polypeptide comprising an amino acid sequence in which at least one amino acids is deleted, replaced, inserted or added in the amino acid sequence of the polypeptide and having substantially the same activity as that of the polypeptide is included in the scope of the present invention. The term “substantially the same activity as that of the polypeptide” means the same activity represented by the inherent function, enzyme activity or the like possessed by the polypeptide which has not been deleted, replaced, inserted or added. The polypeptide can be obtained using a method for introducing part-specific mutation(s) described in, for example, Molecular Cloning, 2nd ed., Current Protocols in Molecular Biology, Nuc. Acids. Res., 10: 6487 (1982), Proc. Natl. Acad. Sci. USA, 79: 6409 (1982), Gene, 34: 315 (1985), Nuc. Acids. Res., 13: 4431 (1985), Proc. Natl. Acad. Sci. USA, 82: 488 (1985) and the like. For example, the polypeptide can be obtained by introducing mutation(s) to DNA encoding a polypeptide having the amino acid sequence represented by any one of SEQ ID NOS:3502 to 6931. The number of the amino acids which are deleted, replaced, inserted or added is not particularly limited; however, it is usually 1 to the order of tens, preferably 1 to 20, more preferably 1 to 10, and most preferably 1 to 5, amino acids.
The at least one amino acid deletion, replacement, insertion or addition in the amino acid sequence of the polypeptide of the present invention is used herein to refer to that at least one amino acid is deleted, replaced, inserted or added to at one or plural positions in the amino acid sequence. The deletion, replacement, insertion or addition may be caused in the same amino acid sequence simultaneously. Also, the amino acid residue replaced, inserted or added can be natural or non-natural. Examples of the natural amino acid residue include L-alanine, L-asparagine, L-asparatic acid, L-glutamine, L-glutamic acid, glycine, L-histidine, L-isoleucine, L-leucine, L-lysine, L-methionine, L-phenylalanine, L-proline, L-serine, L-threonine, L-tryptophan, L-tyrosine, L-valine, L-cysteine, and the like.
Herein, examples of amino acid residues which are replaced with each other are shown below. The amino acid residues in the same group can be replaced with each other.
Group A:
leucine, isoleucine, norleucine, valine, norvaline, alanine, 2-aminobutanoic acid, methionine, O-methylserine, t-butylglycine, t-butylalanine, cyclohexylalanine;
Group B:
asparatic acid, glutamic acid, isoasparatic acid, isoglutamic acid, 2-aminoadipic acid, 2-aminosuberic acid;
Group C:
asparagine, glutamine;
Group D:
lysine, arginine, ornithine, 2,4-diaminobutanoic acid, 2,3-diaminopropionic acid;
Group E:
proline, 3-hydroxyproline, 4-hydroxyproline;
Group F:
serine, threonine, homoserine;
Group G:
phenylalanine, tyrosine.
Also, in order that the resulting mutant polypeptide has substantially the same activity as that of the polypeptide which has not been mutated, it is preferred that the mutant polypeptide has a homology of 60% or more, preferably 80% or more, and particularly preferably 95% or more, with the polypeptide which has not been mutated, when calculated, for example, using default (initial setting) parameters by a homology searching software, such as BLAST, FASTA, or the like.
Also, the polypeptide of the present invention can be produced by a chemical synthesis method, such as Fmoc (fluorenylmethyloxycarbonyl) method, tBoc (t-butyloxycarbonyl) method, or the like. It can also be synthesized using a peptide synthesizer manufactured by Advanced ChemTech, Perkin-Elmer, Pharmacia, Protein Technology Instrument, Synthecell-Vega, PerSeptive, Shimadzu Corporation, or the like.
The transformant of the present invention can be used for objects other than the production of the polypeptide of the present invention.
Specifically, at least one component selected from an amino acid, a nucleic acid, a vitamin, a saccharide, an organic acid, and analogues thereof can be produced by culturing the transformant containing the polynucleotide or recombinant vector of the present invention in a medium to produce and accumulate at least one component selected from amino acids, nucleic acids, vitamins, saccharides, organic acids, and analogues thereof, and recovering the same from the medium.
The biosynthesis pathways, decomposition pathways and regulatory mechanisms of physiologically active substances such as amino acids, nucleic acids, vitamins, saccharides, organic acids and analogues thereof differ from organism to organism. The productivity of such a physiologically active substance can be improved using these differences, specifically by introducing a heterogeneous gene relating to the biosynthesis thereof. For example, the content of lysine, which is one of the essential amino acids, in a plant seed was improved by introducing a synthase gene derived from a bacterium (WO 93/19190). Also, arginine is excessively produced in a culture by introducing an arginine synthase gene derived from Escherichia coli (Japanese Examined Patent Publication 23750/93).
To produce such a physiologically active substance, the transformant according to the present invention can be cultured by the same method as employed in culturing the transformant for producing the polypeptide of the present invention as described above. Also, the physiologically active substance can be recovered from the culture medium in combination with, for example, the ion exchange resin method, the precipitation method and other known methods.
Examples of methods known to one of ordinary skill in the art include electroporation, calcium transfection, the protoplast method, the method using a phage, and the like, when the host is a bacterium; and microinjection, calcium phosphate transfection, the positively charged lipid-mediated method and the method using a virus, and the like, when the host is a eukaryote (Molecular Cloning, 2nd. ed.; Spector et al., Cells/a laboratory manual, Cold Spring Harbour Laboratory Press, 1998)). Examples of the host include prokaryotes, lower eukaryotes (for example, yeasts), higher eukaryotes (for example, mammals), and cells isolated therefrom. As the state of a recombinant polynucleotide fragment present in the host cells, it can be integrated into the chromosome of the host. Alternatively, it can be integrated into a factor (for example, a plasmid) having an independent replication unit outside the chromosome. These transformants are usable in producing the polypeptides of the present invention encoded by the ORF of the genome of Corynebacterium glutamicum, the polynucleotides of the present invention and fragments thereof. Alternatively, they can be used in producing arbitrary polypeptides under the regulation by an EMF of the present invention.
11. Preparation of Antibody Recognizing the Polypeptide of the Present Invention
An antibody which recognizes the polypeptide of the present invention, such as a polyclonal antibody, a monoclonal antibody, or the like, can be produced using, as an antigen, a purified product of the polypeptide of the present invention or a partial fragment polypeptide of the polypeptide or a peptide having a partial amino acid sequence of the polypeptide of the present invention.
(1) Production of Polyclonal Antibody
A polyclonal antibody can be produced using, as an antigen, a purified product of the polypeptide of the present invention, a partial fragment polypeptide of the polypeptide, or a peptide having a partial amino acid sequence of the polypeptide of the present invention, and immunizing an animal with the same.
Examples of the animal to be immunized include rabbits, goats, rats, mice, hamsters, chickens and the like.
A dosage of the antigen is preferably 50 to 100 μg per animal.
When the peptide is used as the antigen, it is preferably a peptide covalently bonded to a carrier protein, such as keyhole limpet haemocyanin, bovine thyroglobulin, or the like. The peptide used as the antigen can be synthesized by a peptide synthesizer.
The administration of the antigen is, for example, carried out 3 to 10 times at the intervals of 1 or 2 weeks after the first administration. On the 3rd to 7th day after each administration, a blood sample is collected from the venous plexus of the eyeground, and it is confirmed that the serum reacts with the antigen by the enzyme immunoassay (Enzyme-linked Immunosorbent Assay (ELISA), Igaku Shoin (1976); Antibodies—A Laboratory Manual, Cold Spring Harbor Laboratory (1988)) or the like.
Serum is obtained from the immunized non-human mammal with a sufficient antibody titer against the antigen used for the immunization, and the serum is isolated and purified to obtain a polyclonal antibody.
Examples of the method for the isolation and purification include centrifugation, salting out by 40-50% saturated ammonium sulfate, caprylic acid precipitation (Antibodies, A Laboratory manual, Cold Spring Harbor Laboratory (1988)), or chromatography using a DEAE-Sepharose column, an anion exchange column, a protein A- or G-column, a gel filtration column, and the like, alone or in combination thereof, by methods known to those of ordinary skill in the art.
(2) Production of Monoclonal Antibody
(a) Preparation of Antibody-Producing Cell
A rat having a serum showing an enough antibody titer against a partial fragment polypeptide of the polypeptide of the present invention used for immunization is used as a supply source of an antibody-producing cell.
On the 3rd to 7th day after the antigen substance is finally administered the rat showing the antibody titer, the spleen is excised.
The spleen is cut to pieces in MEM medium (manufactured by Nissui Pharmaceutical), loosened using a pair of forceps, followed by centrifugation at 1,200 rpm for 5 minutes, and the resulting supernatant is discarded.
The spleen in the precipitated fraction is treated with a Tris-ammonium chloride buffer (pH 7.65) for 1 to 2 minutes to eliminate erythrocytes and washed three times with MEM medium, and the resulting spleen cells are used as antibody-producing cells.
(b) Preparation of Myeloma Cells
As myeloma cells, an established cell line obtained from mouse or rat is used. Examples of useful cell lines include those derived from a mouse, such as P3-X63Ag8-U1 (hereinafter referred to as “UP3-U1”) (Curr. Topics in Microbiol. Immunol., 81: 1 (1978); Europ. J. Immunol., 6: 511 (1976)); SP2/O-Ag14 (SP-2) (Nature, 276: 269 (1978)): P3-X63-Ag8653 (653) (J. Immunol, 123: 1548 (1979)); P3-X63-Ag8 (X63) cell line (Nature, 256: 495 (1975)), and the like, which are 8-azaguanine-resistant mouse (BALB/c) myeloma cell lines. These cell lines are subcultured in 8-azaguanine medium (medium in which, to a medium obtained by adding. 1.5 mmol/l glutamine, 5×10−5 mol/l 2-mercaptoethanol, 10 μg/ml gentamicin and 10% fetal calf serum (FCS) (manufactured by CSL) to RPMI-1640 medium (hereinafter referred to as the “normal medium”), 8-azaguanine is further added at 15 μg/ml) and cultured in the normal medium 3 or 4 days before cell fusion, and 2×107 or more of the cells are used for the fusion.
(c) Production of Hybridoma
The antibody-producing cells obtained in (a) and the myeloma cells obtained in (b) are washed with MEM medium or PBS (disodium hydrogen phosphate: 1.83 g, sodium dihydrogen phosphate: 0.21 g, sodium chloride: 7.65 g, distilled water: 1 liter, pH: 7.2) and mixed to give a ratio of antibody-producing cells:myeloma cells=5:1 to 10:1, followed by centrifugation at 1,200 rpm for 5 minutes, and the supernatant is discarded.
The cells in the resulting precipitated fraction were thoroughly loosened, 0.2 to 1 ml of a mixed solution of 2 g of polyethylene glycol-1000 (PEG-1000), 2 ml of MEM medium and 0.7 ml of dimethylsulfoxide (DMSO) per 108 antibody-producing cells is added to the cells under stirring at 37° C., and then 1 to 2 ml of MEM medium is further added thereto several times at 1 to 2 minute intervals.
After the addition, MEM medium is added to give a total amount of 50 ml. The resulting prepared solution is centrifuged at 900 rpm for 5 minutes, and then the supernatant is discarded. The cells in the resulting precipitated fraction were gently loosened and then gently suspended in 100 ml of HAT medium (the normal medium to which 10−4 mol/l hypoxanthine, 1.5×10−5 mol/l thymidine and 4×10−7 mol/l aminopterin have been added) by repeated drawing up into and discharging from a measuring pipette.
The suspension is poured into a 96 well culture plate at 100 μl/well and cultured at 37° C. for 7 to 14 days in a 5% CO2 incubator.
After culturing, a part of the culture supernatant is recovered, and a hybridoma which specifically reacts with a partial fragment polypeptide of the polypeptide of the present invention is selected according to the enzyme immunoassay described in Antibodies, A Laboratory manual, Cold Spring Harbor Laboratory, Chapter 14 (1998) and the like.
A specific example of the enzyme immunoassay is described below.
The partial fragment polypeptide of the polypeptide of the present invention used as the antigen in the immunization is spread on a suitable plate, is allowed to react with a hybridoma culturing supernatant or a purified antibody obtained in (d) described below as a first antibody, and is further allowed to react with an anti-rat or anti-mouse immunoglobulin antibody labeled with an enzyme, a chemical luminous substance, a radioactive substance or the like as a second antibody for reaction suitable for the labeled substance. A hybridoma which specifically reacts with the polypeptide of the present invention is selected as a hybridoma capable of producing a monoclonal antibody of the present invention.
Cloning is repeated using the hybridoma twice by limiting dilution analysis (HT medium (a medium in which aminopterin has been removed from HAT medium) is firstly used, and the normal medium is secondly used), and a hybridoma which is stable and contains a sufficient amount of antibody titer is selected as a hybridoma capable of producing a monoclonal antibody of the present invention.
(d) Preparation of Monoclonal Antibody
The monoclonal antibody-producing hybridoma cells obtained in (c) are injected intraperitoneally into 8- to 10-week-old mice or nude mice treated with pristane (intraperitoneal administration of 0.5 ml of 2,6,10,14-tetramethylpentadecane (pristane) followed by 2 weeks of feeding) at 5×106 to 20×106 cells/animal. The hybridoma causes ascites tumor in 10 to 21 days.
The ascitic fluid is collected from the mice or nude mice, and centrifuged to remove solid contents at 3000 rpm for 5 minutes.
A monoclonal antibody can be purified and isolated from the resulting supernatant according to the method similar to that used in the polyclonal antibody.
The subclass of the antibody can be determined using a mouse monoclonal antibody typing kit or a rat monoclonal antibody typing kit. The polypeptide amount can be determined by the Lowry method or by calculation based on the absorbance at 280 nm.
The antibody obtained in the above is within the scope of the antibody of the present invention.
The antibody can be used for the general assay using an antibody, such as a radioactive material labeled immunoassay (RIA), competitive binding assay, an immunotissue chemical staining method (ABC method, CSA method, etc.), immunoprecipitation, Western blotting, ELISA assay, and the like (An introduction to Radioimmunoassay and Related Techniques, Elsevier Science (1986); Techniques in Immunocytochemistry, Academic Press, Vol. 1 (1982), Vol. 2 (1983) & Vol. 3 (1985); Practice and Theory of Enzyme Immunoassays, Elsevier Science (1985); Enzyme-linked Immunosorbent Assay (ELISA), Igaku Shoin (1976); Antibodies—A Laboratory Manual, Cold Spring Harbor laboratory (1988); Monoclonal Antibody Experiment Manual, Kodansha Scientific (1987); Second Series Biochemical Experiment Course, Vol. 5, Immunobiochemistry Research Method, Tokyo Kagaku Dojin (1986)).
The antibody of the present invention can be used as it is or after being labeled with a label.
Examples of the label include radioisotope, an affinity label (e.g., biotin, avidin, or the like), an enzyme label (e.g., horseradish peroxidase, alkaline phosphatase, or the like), a fluorescence label (e.g., FITC, rhodamine, or the like), a label using a rhodamine atom, (J. Histochem. Cytochem., 18: 315 (1970); Meth. Enzym., 62: 308 (1979); Immunol., 109: 129 (1972); J. Immunol., Meth., 13: 215 (1979)), and the like.
Expression of the polypeptide of the present invention, fluctuation of the expression, the presence or absence of structural change of the polypeptide, and the presence or absence in an organism other than coryneform bacteria of a polypeptide corresponding to the polypeptide can be analyzed using the antibody or the labeled antibody by the above assay, or a polypeptide array or proteome analysis described below.
Furthermore, the polypeptide recognized by the antibody can be purified by immunoaffinity chromatography using the antibody of the present invention.
12. Production and Use of Polypeptide Array
(1) Production of Polypeptide Array
A polypeptide array can be produced using the polypeptide of the present invention obtained in the above item 10 or the antibody of the present invention obtained in the above item 11.
The polypeptide array of the present invention includes protein chips, and comprises a solid support and the polypeptide or antibody of the present invention adhered to the surface of the solid support.
Examples of the solid support include plastic such as polycarbonate or the like; an acrylic resin, such as polyacrylamide or the like; complex carbohydrates, such as agarose, sepharose, or the like; silica; a silica-based material, carbon, a metal, inorganic glass, latex beads, and the like.
The polypeptides or antibodies according to the present invention can be adhered to the surface of the solid support according to the method described in Biotechniques, 27: 1258-61 (1999); Molecular Medicine Today, 5: 326-7 (1999); Handbook of Experimental Immunology, 4th edition, Blackwell Scientific Publications, Chapter 10 (1986); Meth. Enzym., 34 (1974); Advances in Experimental Medicine and Biology, 42 (1974); U.S. Pat. No. 4,681,870; U.S. Pat. No. 4,282,287; U.S. Pat. No. 4,762,881, or the like.
The analysis described herein can be efficiently performed by adhering the polypeptide or antibody of the present invention to the solid support at a high density, though a high fixation density is not always necessary.
(2) Use of Polypeptide Array
A polypeptide or a compound capable of binding to and interacting with the polypeptides of the present invention adhered to the array can be identified using the polypeptide array to which the polypeptides of the present invention have been adhered thereto as described in the above (1).
Specifically, a polypeptide or a compound capable of binding to and interacting with the polypeptides of the present invention can be identified by subjecting the polypeptides of the present invention to the following steps (i) to (iv):
(i) preparing a polypeptide array having the polypeptide of the present invention adhered thereto by the method of the above (1);
(ii) incubating the polypeptide immobilized on the polypeptide array together with at least one of a second polypeptide or compound;
(iii) detecting any complex formed between the at least one of a second polypeptide or compound and the polypeptide immobilized on the array using, for example, a label bound to the at least one of a second polypeptide or compound, or a secondary label which specifically binds to the complex or to a component of the complex after unbound material has been removed; and
(iv) analyzing the detection data.
Specific examples of the polypeptide array to which the polypeptide of the present invention has been adhered include a polypeptide array containing a solid support to which at least one of a polypeptide containing an amino acid sequence selected from SEQ ID NOS:3502 to 7001, a polypeptide containing an amino acid sequence in which at least one amino acids is deleted, replaced, inserted or added in the amino acid sequence of the polypeptide and having substantially the same activity as that of the polypeptide, a polypeptide containing an amino acid sequence having a homology of 60% or more with the amino acid sequences of the polypeptide and having substantially the same activity as that of the polypeptides, a partial fragment polypeptide, and a peptide comprising an amino acid sequence of a part of a polypeptide.
The amount of production of a polypeptide derived from coryneform bacteria can be analyzed using a polypeptide array to which the antibody of the present invention has been adhered in the above (1).
Specifically, the expression amount of a gene derived from a mutant of coryneform bacteria can be analyzed by subjecting the gene to the following steps (i) to (iv):
(i) preparing a polypeptide array by the method of the above (1);
(ii) incubating the polypeptide array (the first antibody) together with a polypeptide derived from a mutant of coryneform bacteria;
(iii) detecting the polypeptide bound to the polypeptide immobilized on the array using a labeled second antibody of the present invention; and
(iv) analyzing the detection data.
Specific examples of the polypeptide array to which the antibody of the present invention is adhered include a polypeptide array comprising a solid support to which at least one of an antibody which recognizes a polypeptide comprising an amino acid sequence selected from SEQ ID NOS:3502 to 7001, a polypeptide comprising an amino acid sequence in which at least one amino acids is deleted, replaced, inserted or added in the amino acid sequence of the polypeptide and having substantially the same activity as that of the polypeptide, a polypeptide comprising an amino acid sequence having a homology of 60% or more with the amino acid sequences of the polypeptide and having substantially the same activity as that of the polypeptides, a partial fragment polypeptide, or a peptide comprising an amino acid sequence of a part of a polypeptide.
A fluctuation in an expression amount of a specific polypeptide can be monitored using a polypeptide obtained in the time course of culture as the polypeptide derived from coryneform bacteria. The culturing conditions can be optimized by analyzing the fluctuation.
When a polypeptide derived from a mutant of coryneform bacteria is used, a mutated polypeptide can be detected.
13. Identification of Useful Mutation in Mutant by Proteome Analysis
Usually, the proteome is used herein to refer to a method wherein a polypeptide is separated by two-dimensional electrophoresis and the separated polypeptide is digested with an enzyme, followed by identification of the polypeptide using a mass spectrometer (MS) and searching a data base.
The two dimensional electrophoresis means an electrophoretic method which is performed by combining two electrophoretic procedures having different principles. For example, polypeptides are separated depending on molecular weight in the primary electrophoresis. Next, the gel is rotated by 90° or 180° and the secondary electrophoresis is carried out depending on isoelectric point. Thus, various separation patterns can be achieved (JIS K 3600 2474).
In searching the data base, the amino acid sequence information of the polypeptides of the present invention and the recording medium of the present invention provide for in the above items 2 and 8 can be used.
The proteome analysis of a coryneform bacterium and its mutant makes it possible to identify a polypeptide showing a fluctuation therebetween.
The proteome analysis of a wild type strain of coryneform bacteria and a production strain showing an improved productivity of a target product makes it possible to efficiently identify a mutation protein which is useful in breeding for improving the productivity of a target product or a protein of which expression amount is fluctuated.
Specifically, a wild type strain of coryneform bacteria and a lysine-producing strain thereof are each subjected to the proteome analysis. Then, a spot increased in the lysine-producing strain, compared with the wild type strain, is found and a data base is searched so that a polypeptide showing an increase in yield in accordance with an increase in the lysine productivity can be identified. For example, as a result of the proteome analysis on a wild type strain and a lysine-producing strain, the productivity of the catalase having the amino acid sequence represented by SEQ ID NO:3785 is increased in the lysine-producing mutant.
As a result that a protein having a high expression level is identified by proteome analysis using the nucleotide sequence information and the amino acid sequence information, of the genome of the coryneform bacteria of the present invention, and a recording medium storing the sequences, the nucleotide sequence of the gene encoding this protein and the nucleotide sequence in the upstream thereof can be searched at the same time, and thus, a nucleotide sequence having a high expression promoter can be efficiently selected.
In the proteome analysis, a spot on the two-dimentional electrophoresis gel showing a fluctuation is sometimes derived from a modified protein. However, the modified protein can be efficiently identified using the recording medium storing the nucleotide sequence information, the amino acid sequence information, of the genome of coryneform bacteria, and the recording medium storing the sequences, according to the present invention.
Moreover, a useful mutation point in a useful mutant can be easily specified by searching a nucleotide sequence (nucleotide sequence of promoters, ORF, or the like) relating to the thus identified protein using a recording medium storing the nucleotide sequence information and the amino acid sequence information, of the genome of coryneform bacteria of the present invention, and a recording medium storing the sequences and using a primer designed on the basis of the detected nucleotide sequence. As a result that the useful mutation point is specified, an industrially useful mutant having the useful mutation or other useful mutation derived therefrom can be easily bred.
The present invention will be explained in detail below based on Examples. However, the present invention is not limited thereto.
EXAMPLE 1
Determination of the Full Nucleotide Sequence of Genome of Corynebacterium glutamicum
The full nucleotide sequence of the genome of Corynebacterium glutamicum was determined based on the whole genome shotgun method (Science, 269: 496-512 (1995)). In this method, a genome library was prepared and the terminal sequences were determined at random. Subsequently, these sequences were ligated on a computer to cover the full genome. Specifically, the following procedure was carried out.
(1) Preparation of Genome DNA of Corynebacterium glutamicum ATCC 13032
Corynebacterium glutamicum ATCC 13032 was cultured in BY medium (7 g/l meat extract, 10 g/l peptone, 3 g/l sodium chloride, 5 g/l yeast extract, pH 7.2) containing 1% of glycine at 30° C. overnight and the cells were collected by centrifugation. After washing with STE buffer (10.3% sucrose, 25 mmol/l Tris hydrochloride, 25 mmol/l EDTA, pH 8.0), the cells were suspended in 10 ml of STE buffer containing 10 mg/ml lysozyme, followed by gently shaking at 37° C. for 1 hour. Then, 2 ml of 10% SDS was added thereto to lyse the cells, and the resultant mixture was maintained at 65° C. for 10 minutes and then cooled to room temperature. Then, 10 ml of Tris-neutralized phenol was added thereto, followed by gently shaking at room temperature for 30 minutes and centrifugation (15,000×g, 20 minutes, 20° C.). The aqueous layer was separated and subjected to extraction with phenol/chloroform and extraction with chloroform (twice) in the same manner. To the aqueous layer, 3 mol/l sodium acetate solution (pH 5.2) and isopropanol were added at 1/10 times volume and twice volume, respectively, followed by gently stirring to precipitate the genome DNA. The genome DNA was dissolved again in 3 ml of TE buffer (10 mmol/l Tris hydrochloride, 1 mmol/l EDTA, pH 8.0) containing 0.02 mg/ml of RNase and maintained at 37° C. for 45 minutes. The extractions with phenol, phenol/chloroform and chloroform were carried out successively in the same manner as the above. The genome DNA was subjected to isopropanol precipitation. The thus formed genome DNA precipitate was washed with 70% ethanol three times, followed by air-drying, and dissolved in 1.25 ml of TE buffer to give a genome DNA solution (concentration: 0.1 mg/ml).
(2) Construction of a Shotgun Library
TE buffer was added to 0.01 mg of the thus prepared genome DNA of Corynebacterium glutamicum ATCC 13032 to give a total volume of 0.4 ml, and the mixture was treated with a sonicator (Yamato Powersonic Model 150) at an output of 20 continuously for 5 seconds to obtain fragments of 1 to 10 kb. The genome fragments were blunt-ended using a DNA blunting kit (manufactured by Takara Shuzo) and then fractionated by 6% polyacrylamide gel electrophoresis. Genome fragments of 1 to 2 kb were cut out from the gel, and 0.3 ml MG elution buffer (0.5 mol/l ammonium acetate, 10 mmol/l magnesium acetate, 1 mmol/l EDTA, 0.1% SDS) was added thereto, followed by shaking at 37° C. overnight to elute DNA. The DNA eluate was treated with phenol/chloroform, and then precipitated with ethanol to obtain a genome library insert. The total insert and 500 ng of pUC18 SmaI/BAP (manufactured by Amersham Pharmacia Biotech) were ligated at 16° C. for 40 hours.
The ligation product was precipitated with ethanol and dissolved in 0.01 ml of TE buffer. The ligation solution (0.001 ml) was introduced into 0.04 ml of E. coli ELECTRO MAX DH10B (manufactured by Life Technologies) by the electroporation under conditions according to the manufacture's instructions. The mixture was spread on LB plate medium (LB medium (10 g/l bactotrypton, 5 g/l yeast extract, 10 g/l sodium chloride, pH 7.0) containing 1.6% of agar) containing 0.1 mg/ml ampicillin, 0.1 mg/ml X-gal and 1 mmol/l isopropyl-β-D-thiogalactopyranoside (IPTG) and cultured at 37° C. overnight.
The transformant obtained from colonies formed on the plate medium was stationarily cultured in a 96-well titer plate having 0.05 ml of LB medium containing 0.1 mg/ml ampicillin at 37° C. overnight. Then, 0.05 ml of LB medium containing 20% glycerol was added thereto, followed by stirring to obtain a glycerol stock.
(3) Construction of Cosmid Library
About 0.1 mg of the genome DNA of Corynebacterium glutamicum ATCC 13032 was partially digested with Sau3AI (manufactured by Takara Shuzo) and then ultracentrifuged (26,000 rpm, 18 hours, 20° C.) under 10 to 40% sucrose density gradient obtained using 10% and 40% sucrose buffers (1 mol/l NaCl, 20 mmol/l Tris hydrochloride, 5 mmol/l EDTA, 10% or 40% sucrose, pH 8.0). After the centrifugation, the solution thus separated was fractionated into tubes at 1 ml in each tube. After confirming the DNA fragment length of each fraction by agarose gel electrophoresis, a fraction containing a large amount of DNA fragment of about 40 kb was precipitated with ethanol.
The DNA fragment was ligated to the BamHI site of superCos1 (manufactured by Stratagene) in accordance with the manufacture's instructions. The ligation product was incorporated into Escherichia coli XL-1-BlueMR strain (manufactured by Stratagene) using Gigapack III Gold Packaging Extract (manufactured by Stratagene) in accordance with the manufacture's instructions. The Escherichia coli was spread on LB plate medium containing 0.1 mg/ml ampicillin and cultured therein at 37° C. overnight to isolate colonies. The resulting colonies were stationarily cultured at 37° C. overnight in a 96-well titer plate containing 0.05 ml of the LB medium containing 0.1 mg/ml ampicillin in each well. LB medium containing 20% glycerol (0.05 ml) was added thereto, followed by stirring to obtain a glycerol stock.
(4) Determination of Nucleotide Sequence
(4-1) Preparation of Template
The full nucleotide sequence of Corynebacterium glutamicum ATCC 13032 was determined mainly based on the whole genome shotgun method. The template used in the whole genome shotgun method was prepared by the PCR-method using the library prepared in the above (2).
Specifically, the clone derived from the whole genome shotgun library was inoculated using a replicator (manufactured by GENETIX) into each well of a 96-well plate containing the LB medium containing 0.1 mg/ml of ampicillin at 0.08 ml per each well and then stationarily cultured at 37° C. overnight.
Next, the culturing solution was transported using a copy plate (manufactured by Tokken) into a 96-well reaction plate (manufactured by PE Biosystems) containing a PCR reaction solution (TaKaRa Ex Taq (manufactured by Takara Shuzo)) at 0.08 ml per each well. Then, PCR was carried out in accordance with the protocol by Makino et. al. (DNA Research, 5: 1-9 (1998)) using GeneAmp PCR System 9700 (manufactured by PE Biosystems) to amplify the inserted fragment.
The excessive primers and nucleotides were eliminated using a kit for purifying a PCR production (manufactured by Amersham Pharmacia Biotech) and the residue was used as the template in the sequencing reaction.
Some nucleotide sequences were determined using a double-stranded DNA plasmid as a template.
The double-stranded DNA plasmid as the template was obtained by the following method.
The clone derived from the whole genome shotgun library was inoculated into a 24- or 96-well plate containing a 2× YT medium (16 g/l bactotrypton, 10 g/l yeast extract, 5 g/l sodium chloride, pH 7.0) containing 0.05 mg/ml ampicillin at 1.5 ml per each well and then cultured under shaking at 37° C. overnight.
The double-stranded DNA plasmid was prepared from the culturing solution using an automatic plasmid preparing machine, KURABO PI-50 (manufactured by Kurabo Industries) or a multiscreen (manufactured by Millipore) in accordance with the protocol provided by the manufacturer.
To purify the double-stranded DNA plasmid using the multiscreen, Biomek 2000 (manufactured by Beckman Coulter) or the like was employed.
The thus obtained double-stranded DNA plasmid was dissolved in water to give a concentration of about 0.1 mg/ml and used as the template in sequencing.
(4-2) Sequencing Reaction
To 6 μl of a solution of ABI PRISM BigDye Terminator Cycle Sequencing Ready Reaction Kit (manufactured by PE Biosystems), an M13 regular direction primer (M13-21) or an M13 reverse direction primer (M13REV) (DNA Research, 5: 1-9 (1998) and the template prepared in the above (4-1) (the PCR product or the plasmid) were added to give 10 μl of a sequencing reaction solution. The primers and the templates were used in an amount of 1.6 pmol and an amount of 50 to 200 ng, respectively.
Dye terminator sequencing reaction of 45 cycles was carried out with GeneAmp PCR System 9700 (manufactured by PE Biosystems) using the reaction solution. The cycle parameter was determined in accordance with the manufacturer's instruction accompanying ABI PRISM BigDye Terminator Cycle Sequencing Ready Reaction Kit. The sample was purified using MultiScreen HV plate (manufactured by Millipore) according to the manufacture's instructions. The thus purified reaction product was precipitated with ethanol, followed by drying, and then stored in the dark at −30° C.
The dry reaction product was analyzed by ABI PRISM 377 DNA Sequencer and ABI PRISM 3700 DNA Analyzer (both manufactured by PE Biosystems) each in accordance with the manufacture's instructions.
The data of about 50,000 sequences in total (i.e., about 42,000 sequences obtained using 377 DNA Sequencer and about 8,000 reactions obtained by 3700 DNA Analyser) were transferred to a server (Alpha Server 4100: manufactured by COMPAQ) and stored. The data of these about 50,000 sequences corresponded to 6 times as much as the genome size.
(5) Assembly
All operations were carried out on the basis of UNIX platform. The analytical data were output in Macintosh platform using X Window System. The base call was carried out using phred (The University of Washington). The vector sequence data was deleted using SPS Cross_Match (manufactured by Southwest Parallel Software). The assembly was carried out using SPS phrap (manufactured by Southwest Parallel Software; a high-speed version of phrap (The University of Washington)). The contig obtained by the assembly was analyzed using a graphical editor, consed (The University of Washington). A series of the operations from the base call to the assembly were carried out simultaneously using a script phredPhrap attached to consed.
(6) Determination of Nucleotide Sequence in Gap Part
Each cosmid in the cosmid library constructed in the above (3) was prepared by a method similar to the preparation of the double-stranded DNA plasmid described in the above (4-1). The nucleotide sequence at the end of the inserted fragment of the cosmid was determined by using ABI PRISM BigDye Terminator Cycle Sequencing Ready Reaction Kit (manufactured by PE Biosystems) according to the manufacture's instructions.
About 800 cosmid clones were sequenced at both ends to search a nucleotide sequence in the contig derived from the shotgun sequencing obtained in the above (5) coincident with the sequence. Thus, the linkage between respective cosmid clones and respective contigs were determined and mutual alignment was carried out. Furthermore, the results were compared with the physical map of Corynebacterium glutamicum ATCC 13032 (Mol. Gen. Genet., 252: 255-265 (1996) to carrying out mapping between the cosmids and the contigs.
The sequence in the region which was not covered with the contigs was determined by the following method.
Clones containing sequences positioned at the ends of contigs were selected. Among these clones, about 1,000 clones wherein only one end of the inserted fragment had been determined were selected and the sequence at the opposite end of the inserted fragment was determined. A shotgun library clone or a cosmid clone containing the sequences at the respective ends of the inserted fragment in two contigs was identified, the full nucleotide sequence of the inserted fragment of this clone was determined, and thus the nucleotide sequence of the gap part was determined. When no shotgun library clone or cosmid clone covering the gap part was available, primers complementary to the end sequences at the two contigs were prepared and the DNA fragment in the gap part was amplified by PCR. Then, sequencing was performed by the primer walking method using the amplified DNA fragment as a template or by the shotgun method in which the sequence of a shotgun clone prepared from the amplified DNA fragment was determined. Thus, the nucleotide sequence of the domain was determined.
In a region showing a low sequence precision, primers were synthesized using AUTOFINISH function and NAVIGATING function of consed (The University of Washington) and the sequence was determined by the primer walking method to improve the sequence precision. The thus determined full nucleotide sequence of the genome of Corynebacterium glutamicum ATCC 13032 strain is shown in SEQ ID NO:1.
(7) Identification of ORF and Presumption of its Function
ORFs in the nucleotide sequence represented by SEQ ID NO:1 were identified according to the following method. First, the ORF regions were determined using software for identifying ORF, i.e., Glimmer, GeneMark and GeneMark.hmm on UNIX platform according to the respective manual attached to the software.
Based on the data thus obtained, ORFs in the nucleotide sequence represented by SEQ ID NO:1 were identified.
The putative function of an ORF was determined by searching the homology of the identified amino acid sequence of the ORF against an amino acid database consisting of protein-encoding domains derived from Swiss-Prot, PIR or Genpept database constituted by protein encoding domains derived from GenBank database, Frame Search (manufactured by Compugen), or by searching the homology of the identified amino acid sequence of the ORF against an amino acid database consisting of protein-encoding domains derived from Swiss-Prot, PIR or Genpept database constituted by protein encoding domains derived from GenBank database, BLAST. The nucleotide sequences of the thus determined ORFs are shown in SEQ ID NOS:2 to 3501, and the amino acid sequences encoded by these ORFs are shown in SEQ ID NOS:3502 to 7001.
In some cases of the sequence listings in the present invention, nucleotide sequences, such as TTG, TGT, GGT, and the like, other than ATG, are read as an initiating codon encoding Met.
Also, the preferred nucleotide sequences are SEQ ID NOS:2 to 355 and 357 to 3501, and the preferred amino acid sequences are shown in SEQ ID NOS:3502 to 3855 and 3957 to 7001
Table 1 shows the registration numbers in the above-described databases of sequences which were judged as having the highest homology with the nucleotide sequences of the ORFs as the results of the homology search in the amino acid sequences using the homology-searching software Frame Search (manufactured by Compugen), names of the genes of these sequences, the functions of the genes, and the matched length, identities and analogies compared with publicly known amino acid translation sequences. Moreover, the corresponding positions were confirmed via the alignment of the nucleotide sequence of an arbitrary ORF with the nucleotide sequence of SEQ ID NO:1. Also, the positions of nucleotide sequences other than the ORFs (for example, ribosomal RNA genes, transfer RNA genes, IS sequences, and the like) on, the genome were determined.
FIG. 1 shows the positions of typical genes of the Corynebacterium glutamicum ATCC 13032 on the genome.
TABLE 1
|
|
TerminalIdentitySimilarityMatched length
SEQ NO. (DNA)SEQ NO. (a.a.)Initial (nt)(nt)ORF (bp)db MatchHomologous gene(%)(%)(a.a.)Function
|
|
23502115721572gsp: R98523Brevibacterium flavum dnaA99.899.8524replication initiation protein DnaA
3350319201597324
43504229234731182sp: DP3B_MYCSMMycobacterium smegmatis dnaN50.581.8390DNA polymerase III beta chain
53505358547661182sp: RECF_MYCSMMycobacterium smegmatis recF53.379.9392DNA replication protein (recF
protein)
6350647665299534sp: YREG_STRCOStreptomyces coelicolor yreG35.158.1174hypothetical protein
73507535474862133pir: S44198Mycobacterium tuberculosis71.988.9704DNA topoisomerase (ATP-
H37Rv gyrBhydrolyzing)
8350878308795966
9350994668798669
103510956210071510
11351199149474441
12351211177101071071sp: YV11_MYCTUMycobacterium tuberculosis29.450.7422NAGC/XYLR repressor
H37Rv
1335131152311263261
1435141176811523246
15351511831143982568sp: GYRA_MYCTUMycobacterium tuberculosis70.488.1854DNA gyrase subunit A
H37Rv Rv0006 gyrA
1635161440514746342pir: E70698Mycobacterium tuberculosis29.569.6112hypothetical membrane protein
H37Rv Rv0007
17351716243152091035sp: YEIH_ECOLIEscherichia coli K12 yeiH33.763.5329hypothetical protein
1835181631417207894gp: AB042619_1Hydrogenophilus thermoluteolus27.662.3268bacterial regulatory protein, LysR
TH-1 cbbRtype
1935191725117670420
2035201872917860870gp: AF156103_2Rhodobacter capsulatus ccdA29.157.4265cytochrome c biogenesis protein
2135211949718736762pir: A49232Coxiella burnetii com131.664.5155hypothetical protein
2235221970520073369pir: F70664Mycobacterium tuberculosis36.870.1117repressor
H37Rv Rv1846c
2335232007321065993gp: MLCB1788_6Mycobacterium leprae24.950.8321hypothetical membrane protein
MLCB1788.18
2435242125321074180pir: I40838Corynebacterium sp. ATCC65.488.5262,5-diketo-D-gluconic acid reductase
31090
2535252159722124528sp: 5NTD_VIBPAVibrio parahaemolyticus nutA27.056.11965′-nucleotidase precursor
26352622164233991236gp: AE001909_7Deinococcus radiodurans27.056.72705′-nucleotidase family protein
DR0505
2735272377923615165prf: 2513302CCorynebacterium striatum ORF152.972.651transposase
2835282429524729435prf: 2413353AXanthomonas campestris51.879.9139organic hydroperoxide detoxication
phaseoli ohrenzyme
29352926297248851413sp: RECG_THIFEThiobacillus ferrooxidans recG32.760.8217ATP-dependent DNA helicase
3035302633826775438
31353128099268221278sp: AMYH_YEASTSaccharomyces cerevisiae26.754.1449glucan 1,4-alpha-glucosidase
S288C YIR019C sta1
3235322911728164954gp: ERU52850_1Erysipelothrix rhusiopathiae28.963.7311lipoprotein
ewlA
3335332996529117849gp: AF180520_3Streptococcus pyogenes SF37034.674.1266ABC 3 transport family or integral
mtsCmembrane protein
3435342999530651657sp: FECE_ECOLIEscherichia coli K12 fecE39.270.3222iron(III) dicitrate transport ATP-
biding protein
3535353069731677981pir: A72417Thermotoga maritima MSB825.856.5283sugar ABC transporter, periplasmic
TM0114sugar-binding protein
36353631677326991023prf: 1207243BEscherichia coli K12 rbsC30.568.3312high affinity ribose transport protein
3735373269933457759sp: RBSA_BACSUBacillus subtilis 168 rbsA32.276.7236ribose transport ATP-binding protein
3835383428033465816pir: I51116Petromyzon marinus23.644.4347neurofilament subunit NF-180
3935393433934899561sp: CYPA_MYCTUMycobacterium leprae H37RV79.989.9169peptidyl-prolyl cls-trans Isomerase A
RV0009 ppiA
4035403498235668687sp: YQGP_BACSUBacillus subtilis 168 yqgP29.253.1226hypothetical membrane protein
4135413722138198978sp: FEPG_ECOLIEscherichia coli K12 fepG40.470.5332ferric enterobactin transport system
permease protein
4235423724236247996
4335433820238978777gp: VCU52150_9Vibrio cholerae viuC51.881.8253ATPase
4435443897839799822sp: VIUB_VIBVUVibrio vulnificus MO6-24 viuB26.252.7260vulnibactin utilization protein
4535454045840189270sp: YO11_MYCTUMycobacterium tuberculosis40.072.695hypothetical membrane protein
H37Rv Rv0011c
46354642513405761938sp: PKNB_MYCLEMycobacterium leprae pknB40.668.7648serine/threonine protein kinase
47354743919425131407gp: AF094711_1Streptomyces coelicolor pksC31.759.1486serine/threonine protein kinase
48354845347439261422gp: AF241575_1Streptomyces griseus pbpA33.566.7492penicillin-binding protein
49354946489453471143sp: SP5E_BACSUBacillus subtilis 168 spoVE31.265.6375stage V sporulation protein E
50355048021466691353pir: H70699Mycobacterium tuberculosis44.170.8469phosphoprotein phosphatase
H37Rv ppp
5135514848548024462pir: A70700Mycobacterium tuberculosis38.766.5155hypothetical protein
H37Rv Rv0019c
5235524936848505864pir: B70700Mycobacterium tuberculosis23.638.8526hypothetical protein
H37Rv Rv0020c
5335534960149455147
5435545061649897720
5535555097250754219
5635565143650966471
5735575305554008954sp: PH2M_TRICUTrichosporon cutaneum ATCC29.963.3117phenol 2-monooxygenase
46490
58355853095516261470sp: GABD_ECOLIEscherichia coli K12 gabD46.778.2490succinate-semialdehyde
dehydrogenase (NAD(P)+)
59355954080555461467sp: YRKH_BACSUBacillus subtilis yrkH27.357.0242hypothetical protein
6035605641755629789sp: Y441_METJAMethanococcus jannaschii29.064.1262hypothetical membrane protein
MJ0441
6135615667656386291sp: YRKF_BACSUBacillus subtilis yrkF40.574.374hypothetical protein
6235625727056680591sp: YC61_SYNY3Synechocystis sp. PCC680336.370.4179hypothetical protein
slr1261
6335635747857651174pir: G70988Mycobacterium tuberculosis53.283.962hypothetical protein
H37Rv Rv1766
6435645808758941855
6535655909159930840gp: LMFL4768_11Leishmania major L4768.1126.850.7310hypothetical protein
6635665995260662711
67356760669623211653
68356863508623901119pir: F70952Mycobacterium tuberculosis29.559.5390magnesium and cobalt transport
H37Rv Rv1239c corAprotein
6935696404063594447
70357064190654581269gp: AF179611_12Zymomonas mobilis ZM4 clcb30.064.8400chloride channel protein
7135716619765508690sp: PNUC_SALTYSalmonella typhimurium pnuC24.153.1241required for NMN transport
72357266851679721122sp: PHOL_MYCTUMycobacterium tuberculosis29.160.0340phosphate starvation-induced
H37Rv RV2368Cprotein-like protein
7335736817068301132
7435746863468251384
7535756906069824765
76357670186687201467sp: CITM_BACSUBacillus subtilis citM42.368.8497Mg(2+)/citrate complex secondary
transporter
77357770506721581653sp: DPIB_ECOLIEscherichia coli K12 dpiB27.260.6563two-component system sensor
histidine kinase
7835787204371474570
7935797216172814654sp: DPIA_ECOLIEscherichia coli K12 criR33.263.3229transcriptional regulator
8035807372872817912gp: AF134895_1Corynebacterium glutamicum43.373.7293D-isomer specific 2-hydroxyacid
unkdhdehydrogenase
8135817384474272429gp: SCM2_3Streptomyces coelicolor A3(2)38.676.4127hypothetical protein
SCM2.03
82358274490754911002sp: BIOB_CORGLCorynebacterium glutamicum99.499.7334biotin synthase
bioB
8335837550675742237pir: H70542Mycobacterium tuberculosis72.179.143hypothetical protein
H37Rv Rv1590
8435847569776035339sp: YKI4_YEASTSaccharomyces cerevisiae34.163.585hypothetical protein
YKL084w
8535857635376469117
8635868075380613141PIR: F81737Chlamydia muridarum Nigg71.075.042hypothetical protein
TC0129
8735878127481002273GSP: Y35814Chlamydia pneumoniae61.066.084hypothetical protein
88358883568821201449prf: 2512333AStreptomyces virginiae varS25.659.0507Integral membrane efflux protein
89358984935836911245gp: D38505_1Bacillus sp.97.299.8394creatinine deaminase
9035908540385098306
9135918627785663615
9235928631887241924sp: HST2_YEASTSaccharomyces cerevisiae hst226.250.2279SIR2 gene family (silent information
regulator)
9335938853287561972prf: 2316378APropionibacterium acnes30.759.0251triacylglycerol lipase
9435948944488545900prf: 2316378APropionibacterium acnes29.456.1262triacylglycerol lipase
9535958955890445888
9635969097390461513gp: AB029154_1Corynebacterium glutamicum90.694.7171transcriptional regulator
ureR
9735979117491473300gp: AB029154_2Corynebacterium glutamicum100.0100.0100urease gammma subunit or urease
ureAstructural protein
9835989150391988486gp: CGL251883_2Corynebacterium glutamicum100.0100.0162urease beta subunit
ATCC 13032 ureB
99359991992937011710gp: CGL251883_3Corynebacterium glutamicum100.0100.0570urease alpha subunit
ATCC 13032 ureC
10036009372994199471gp: CGL251883_4Corynebacterium glutamicum100.0100.0157urease accessory protein
ATCC 13032 ureE
10136019420294879678gp: CGL251883_5Corynebacterium glutamicum100.0100.0226urease accessory protein
ATCC 13032 ureF
10236029489995513615gp: CGL251883_6Corynebacterium glutamicum100.0100.0205urease accessory protein
ATCC 13032 ureG
10336039551796365849gp: CGL251883_7Corynebacterium glutamicum100.0100.0283urease accessory protein
ATCC 13032 ureD
10436049714496368777prf: 2318326BAgrobacterium radiobacter echA21.248.4279epoxide hydrolase
10536059752198189669
106360698470973191152gp: AF148322_1Streptomyces viridifaciens vlmF26.559.7347valanimycin resistant protein
107360799819100493675
1083608101582988082775
10936091034351016121824sp: HTPG_ECOLIEscherichia coli K12 htpG23.852.7668heat shock protein (hsp90-family)
11036101034941049091416sp: AMN_ECOLIEscherichia coli K12 amn41.068.2481AMP nucleosidase
1113611105751105173579
1123612106392105841552pir: E72483Aeropyrum pernix K1 APE250929.658.7196acetolactate synthase large subunit
1133613107289106630660
11436141074351108903456sp: PUTA_SALTYSalmonella typhimurium putA25.850.41297proline dehydrogenase/P5C
dehydrogenase
1153615111161111274114
1163616111374112318945sp: AAD_PHACHPhanerochaete chrysosporium30.260.7338aryl-alcohol dehydrogenase
aad(NADP+)
11736171124701140831614sp: YDAH_ECOLIEscherichia coli K12 ydaH36.571.4513pump protein (transport)
11836181141471154781332prf: 2422424AEnterobacter agglomerans23.049.2352Indole-3-acetyl-Asp hydrolase
1193619115262114564699
1203620115578115943366sp: YIDH_ECOLIEscherichia coli K12 yidH35.970.8106hypothetical membrane protein
1213621115949116263315
12236221185991165482052
1233623119589118810780sp: ACCR_AGRTUAgrobacterium tumefaciens29.559.7258transcriptional repressor
accR
1243624120021120410390pir: C70019Bacillus subtilis yurT57.978.6126methylglyoxalase
1253625120922120413510sp: YC76_MYCTUMycobacterium tuberculosis37.064.8162hypothetical protein
H37Rv Rv1276c
12636261224591209511509prf: 2309180APseudomonas fluorescens mtlD43.570.4497mannitol dehydrogenase
12736271238411225071335prf: 2321326AKlebsiella pneumoniae dalT30.368.3435D-arabinitol transporter
1283628123842124030189
1293629124130124966837sp: GATR_ECOLIEscherichia coli K12 gatR27.364.6260galactitol utilization operon repressor
13036301249321263501419sp: XYLB_STRRUStreptomyces rubiginosus xylB45.068.1451xylulose kinase
1313631127171127992822
1323632127189126353837gp: CGPAN_2Corynebacterium glutamicum100.0100.0279pantoate-beta-alanine ligase
ATCC 13032 panC
1333633128004127192813gp: CGPAN_1Corynebacterium glutamicum100.0100.02713-methyl-2-oxobutanoate
ATCC 13032 panBhydroxymethyltransferase
1343634129049128099951
1353635130118129489630sp: 3MG_ARATHArabidopsis thaliana mag42.067.6188DNA-3-methyladenine glycosylase
1363636130145130798654
1373637131738130815924gp: AB029896_1Petroleum-degrading bacterium39.369.3270esterase
HD-1 hde
1383638131798132424627
1393639132424132981558sp: CAH_METTEMethanosarcina thermophila30.953.2201carbonate dehydratase
14036401341131329711143sp: XYLR_BACSUBacillus subtilis W23 xylR24.149.3357xylose operon repressor protein
14136411354781342071272gp: LLLPK214_12Lactococcus lactis mef21421.161.2418macrolide efflux protein
1423642136321135518804
1433643136565136122444
14436441368041387441941
14536451387911403291539
1463646139861139226636
14736471403291417891461pir: I39714Agrobacterium tumefaciens celA24.351.2420cellulose synthase
14836481417961435261731sp: HKR1_YEASTSaccharomyces cerevisiae25.151.8593hypothetical membrane protein
YDR420W hkr1
1493649142455143075621
15036501435751446391065
1513651144725145480756
1523652146396145518879sp: RARD_PSEAEPseudomonas aeruginosa rarD34.760.7303chloramphenicol sensitive protein
1533653146522147238717sp: YADS_ECOLIEscherichia coli K12 yadS30.359.1198hypothetical membrane protein
1543654147238147570333
15536551481221497801659
15636561509301497941137sp: ABRB_ECOLIEscherichia coli K12 abrB32.462.3361transport protein
1573657151572152369798sp: YFCA_ECOLIEscherichia coli K12 yfcA34.770.2248hypothetical membrane protein
1583658151589150966624
1593659152410152814405
16036601556131532262388sp: HRPB_ECOLIEscherichia coli K12 hrpB33.864.3829ATP-dependent helicase
1613661155853156167315
1623662156821156147675sp: NODL_RHILVRhizobium leguminosarum bv.40.466.0188nodulation protein
viciae plasmid pRL1JI nodL
1633663156848157537690sp: ALKB_ECOLIEscherichia coli o373#1 alkB34.760.7219DNA repair system specific for
alkylated DNA
1643664157614158138525sp: 3MG1_ECOLIEscherichia coli K12 tag39.865.1166DNA-3-methyladenine glycosylase
1653665158154158831678sp: RHTC_ECOLIEscherichia coli K12 rhtC34.161.3217threonine efflux protein
1663666158869159159291sp: YAAA_BACSUBacillus subtilis yaaA50.972.755hypothetical protein
1673667159162160013852prf: 2510326BStreptomyces peucetius dnrV31.052.1284doxorubicin biosynthesis enzyme
1683668160029160370342gp: SPAC1250_3Schizosaccharomyces pombe35.656.7104methyltransferase
SPAC1250.04c
1693669160431161360930
1703670161696162352657
1713671162295161363933
1723672162463162867405gp: AE002420_13Neisseria meningitidis MC5841.576.3118ribonuclease
NMB0662
1733673162965163603639
1743674165717166457741
17536751657551636892067gp: AF176569_1Mus musculus nl128.557.2722neprilysin-like metallopeptidase 1
1763676166457167419963
1773677168595167837759sp: FARR_ECOLIEscherichia coli K12 farR29.865.6238transcriptional regulator, GntR family
or fatty acyl-responsive regulator
17836781689751699911017pir: T14544Beta vulgaris28.663.0332fructokinase or carbohydrate kinase
1793679169996170916921gp: SC8F11_3Streptomyces coelicolor A3(2)52.780.7296hypothetical protein
SC8F11.03c
18036801709331724441512prf: 2204281AStreptomyces coelicolor msdA61.086.1498methylmalonic acid semialdehyde
dehydrogenase
1813681172468173355888sp: IOLB_BACSUBacillus subtilis iolB33.258.2268myo-inositol catabolism
18236821735481752751728sp: IOLD_BACSUBacillus subtilis iolD41.069.8586myo-inositol catabolism
1833683175319176272954sp: MOCC_RHIMERhizobium meliloti mocC29.751.0290rhizopine catabolism protein
18436841763081773181011sp: MI2D_BACSUBacillus subtilis idh or iolG39.172.2335myo-inositol 2-dehydrogenase
1853685177334178203870sp: IOLH_BACSUBacillus subtilis iolH44.672.1287myo-inositol catabolism
18636861782851796581374sp: TCMA_STRGAStreptomyces glaucescens tcmA30.961.5457metabolite export pump of
tetracenomycin C resistance
1873687179081178461621
18836881796891807111023sp: YVAA_BACSUBacillus subtilis yvaA31.165.5354oxidoreductase
1893689180842181297456
1903690181264181647384
1913691182679181687993gp: SRE9798_1Streptomyces reticuli cebR32.061.9331regulatory protein
19236921828191840511233sp: Y4HM_RHISNRhizobium sp. NGR234 y4hM24.452.5442oxidoreductase
19336931840771850871011sp: YFIH_BACSUBacillus subtilis yfiH33.764.7303hypothetical protein
1943694185214185642429
1953695186508186708201sp: CSP_ARTGOStreptomyces coelicolor A3(2)70.392.264cold shock protein
csp
1963696186769187302534
1973697187302187607306
1983698187687188100414prf: 2113413AStellaria longipes30.658.2134caffeoyl-CoA 3-O-methyltransferase
1993699188725188300426
2003700189736188747990sp: CCPA_BACSUBacillus subtilis ccpA28.762.1338glucose-resistance amylase
regulator regulator
2013701189920190321402
2023702190628190389240
20337031921751907031473sp: XYLT_LACBRLactobacillus brevis xylT36.070.5458D-xylose proton symporter
2043704193248192949300
20537051932621944641203gp: AF189147_1Corynebacterium glutamicum100.0100.0401transposase (ISCg2)
ATCC 13032 tnp
2063706195038194604435sp: FIXL_RHIMERhizobium meliloti fixL27.660.7145signal-transducing histidine kinase
20737071952401997694530gp: AB024708_1Corynebacterium glutamicum99.9100.01510glutamine 2-oxoglutarate
gltBaminotransferase large subunit
20837081997722012891518gp: AB024708_2Corynebacterium glutamicum99.499.8506glutamine 2-oxoglutarate
gltDaminotransferase small subunit
2093709201580201341240
21037102032442017601485pir: C70793Mycobacterium tuberculosis44.672.8496hypothetical protein
H37Rv Rv3698
2113711205588205956369
2123712206068206385318
21337132070112035413471prf: 2224383CMycobacterium avium embB39.870.61122arabinosyl transferase
21437142089892070071983pir: D70697Mycobacterium tuberculosis35.066.1651hypothetical membrane protein
H37Rv Rv3792
2153715209968209210759prf: 2504279BPseudomonas sp. phbB31.456.5223acetoacetyl CoA reductase
21637162114552099921464pir: B70697Mycobacterium tuberculosis66.085.1464oxidoreductase
H37Rv Rv3790
2173717211768211535234
2183718211777212283507
2193719212283212735453
22037202126562136571002gp: LMA243459_1Leishmania major ppg124.357.4350proteophosphoglycan
2213721213712214107396sp: Y0GN_MYCTUMycobacterium tuberculosis60.583.9124hypothetical protein
H37Rv Rv3789
2223722214121214522402
2233723214527215159633pir: H70666Mycobacterium tuberculosis43.273.8206hypothetical protein
H37Rv Rv1864c
2243724216100215162939pir: B70696Mycobacterium tuberculosis63.679.1302rhamnosyl transferase
H37Rv Rv3782 rfbE
2253725216264216605342
2263726216712216116597gp: AB016260_100Agrobacterium tumefaciens31.355.1214hypothetical protein
plasmid pTi-SAKURA tiorf100
2273727217929217141789sp: RFBE_YERENYersinia enterocolitica rfbE47.078.4236O-antigen export system ATP-
binding protein
2283728218746217943804sp: RFBD_YERENYersinia enterocolitica rfbD31.375.6262O-antigen export system permease
protein
22937292189792201511173pir: F70695Mycobacterium tuberculosis36.563.0416hypothetical protein
H37Rv Rv3778c
2303730221107220154954gp: AF010309_1Homo sapiens pig341.171.5302NADPH quinone oxidoreductase
2313731221712221131582
2323732221911222207297PIR: A70606Mycobacterium tuberculosis35.051.078probable electron transfer protein
H37Rv Rv3571
23337332236852222101476sp: ALST_BACSUBacillus subtilis alsT46.775.8475amino acid carrier protein
2343734224336225244909
23537352263242252421083gp: SYPCCMOEB_1Synechococcus sp. PCC 794243.870.1368molybdopterin biosynthesis protein
moeBmoeB (sulfurylase)
2363736226767226312456prf: 2403296DArthrobacter nicotinovorans44.775.3150molybdopterin synthase, large
moaEsubunit
2373737227230226760471sp: MOCB_SYNP7Synechococcus sp. PCC 794233.563.3158molybdenum cofactor biosynthesis
moaCBprotein CB
2383738227685227218468prf: 2403296CArthrobacter nicotinovorans61.784.4154co-factor synthesis protein
moaC
23937392288872277031185gp: ANY10817_2Arthrobacter nicotinovorans34.558.6377molybdopterin co-factor synthesis
moeAprotein
2403740229613228891723prf: 2403296FArthrobacter nicotinovorans44.170.5227hypothetical membrane protein
modB
2413741230514229711804prf: 2403296EArthrobacter nicotinovorans34.068.0256molybdate-binding periplasmic
modAprotein
2423742230608230928321pir: D70816Mycobacterium tuberculosis37.570.896molybdopterin converting factor
H37Rv moaD2subunit 1
2433743231842230931912prf: 2518354AThermococcus litoralis malK34.360.8365maltose transport protein
2443744232267231848420sp: YPT3_STRCOStreptomyces coelicolor A3(2)36.476.9121hypothetical membrane protein
ORF3
24537452332822322601023sp: HIS8_ZYMMOZymomonas mobilis hisC37.365.8330histidinol-phosphate
aminotransferase
2463746233913234818906
2473747235203234910294
2483748235290235409120
2493749236212235451762gp: BAU81286_1Brucella abortus oxyR29.457.1252transcription factor
25037502363262373421017sp: ADH2_BACSTBacillius stearothermophilus34.066.0335alcohol dehydrogenase
DSM 2334 adh
2513751237345238145801sp: PUO_MICRUMicrococcus rubens puo21.538.1451putrescine oxidase
25237522381762395251350prf: 2305239ABorrelia burgdorferi mgtE30.968.5444magnesium ion transporter
2533753239772239945174
25437542399862415151530prf: 2320140AXenopus laevis33.259.6567Na/dicarboxylate cotransporter
25537552429022418831020pir: C70800Mycobacterium tuberculosis46.169.1317oxidoreductase
H37Rv tyrA
2563756242910243431522pir: B70800Mycobacterium tuberculosis48.873.8160hypothetical protein
H37Rv Rv3753c
2573757243494243910417gp: RHBNFXP_1Bradyrhizobium japonicum45.170.1144nitrogen fixation protein
2583758244015244215201
2593759244466244816351
26037602449022473042403sp: YV34_MYCTUMycobacterium tuberculosis20.745.7997membrane transport protein
H37Rv Rv0507 mmpL2
26137612473102485721263sp: TGT_ZYMMOZymomonas mobilis41.368.0400queuine tRNA-ribosyltransferase
2623762249294248557738sp: YPDP_BACSUBacillus subtilis ypdP28.162.1203hypothetical membrane protein
26337632494282505071080
2643764250369249722648
26537652505032519391437pir: S65588Streptomyces glaucescens strW24.349.6526ABC transporter
2663766251952252830879sp: SYE_BACSUBacillus subtilis gltX34.863.3316glutamyl-tRNA synthetase
2673767253819252830990
26837682554382543291110gp: PSESTBCBAD_1Pseudomonas syringae tnpA34.255.0360transposase
2693769255794255492303
2703770256067256204138
27137712565992578941296gsp: W69554Brevibacterium lactofermentum98.6100.0432aspartate transaminase
aspC
2723772257900258529630
27337732585512608752325gp: AF025391_1Thermus thermophilus dnaX31.653.1642DNA polymerase III holoenzyme tau
subunit
2743774259312258596717
2753775260987261295309sp: YAAK_BACSUBacillus subtilis yaaK41.674.3101hypothetical protein
2763776261402262055654sp: RECR_BACSUBacillus subtilis recR42.572.4214recombination protein
2773777263295262546750prf: 2503462BHeliobacillus mobilis cobQ38.361.7248cobyric acid synthase
27837782645662632981269prf: 2503462CHeliobacillus mobilis murC31.360.6444UDP-N-acetylmuramyl tripeptide
synthetase
27937792656782645991080pir: H70794Mycobacterium tuberculosis25.755.2346DNA polymerase III epsilon chain
H37Rv dnaQ
2803780269124268258867sp: YLEU_CORGLCorynebacterium glutamicum100.0100.0270hypothetical membrane protein
(Brevibacterium flavum) ATCC
13032 orfX
28137812693712706331263sp: AKAB_CORGLCorynebacterium glutamicum99.599.8421aspartate kinase alpha chain
lysC-alpha
28237822705762695241053
28337832717612731941434
2843784274120273542579prf: 2312309AMycobacterium smegmatis sigE31.263.5189extracytoplasmic function alternative
sigma factor
28537852743662758711506sp: CATV_BACSUBacillus subtilis katA52.976.4492vegetative catalase
2863786275891276232342
2873787276247275957291
2883788276763276302462sp: LRP_KLEPNKlebsiella pneumoniae lrp37.172.0143leucine-responsive regulatory
protein
2893789276829277581753sp: AZLC_BACSUBacillus subtilis 1A1 aziC30.568.0203branched-chain amino acid transport
2903790277581277904324
2913791278301277987315
2923792278732278388345gp: AF178758_1SinoRhizobium sp. As4 arsR34.468.990metalloregulatory protein
29337932788142798931080gp: AF178758_2Sinorhizobium sp. As4 arsB52.284.2341arsenic oxyanion-translocation pump
membrane subunit
2943794279893280279387sp: ARSC_STAXYStaphylococcus xylosus arsC31.168.9119arsenate reductase
2953795280666280349318
2963796280939280670270
2973797281401280949453
29837982829332814041530gp: AF097740_4Bacillus firmus OF4 mrpD32.470.4503Na+/H+ antiporter or multiple
resistance and pH regulation related
protein D
2993799283317282937381prf: 2504285DStaphylococcus aureus mnhC37.070.6119Na+/H+ antiporter
30038002862022833172886gp: AF097740_1Bacillus firmus OF4 mrpA34.164.3824Na+/H+ antiporter or multiple
resistance and pH regulation related
protein A
30138012863732878571485
3023802287661287059603
3033803288829287966864
3043804289796289131666sp: CZCR_ALCEUAlcaligenes eutrophus CH3438.670.4223transcriptional activator
czcR
30538052912432897771467prf: 2214304BMycobacterium tuberculosis26.756.8521two-component system sensor
mtrBhistidine kinase
3063806291815292417603sp: APL_LACLALactococcus lactis MG1363 apl28.360.0180alkaline phosphatase
3073807291833291273561
3083808293511292597915pir: B69865Bacillus subtilis ykuE26.154.7307phosphoesterase
3093809293539293991453sp: YQEY_BACSUBacillus subtilis yqeY37.671.8149hypothetical protein
31038102963882940042385prf: 2209359AMycobacterium leprae pon148.377.1782class A penicillin-binding
protein(PBP1)
3113811297064297402339pir: S20912Streptomyces coelicolor A3(2)40.963.471regulatory protein
whiB
3123812297431297622192
3133813297631297783153gp: SCH17_10Streptomyces coelicolor A3(2)84.096.050hypothetical protein
SCH17.10c
3143814297792298250459pir: G70790Mycobacterium tuberculosis65.189.9149transcriptional regulator
H37Rv Rv3678c
31538152996842983321353sp: SHIA_ECOLIEscherichia coli K12 shiA37.368.9440shikimate transport protein
3163816300087300695609
31738173012612997261536sp: LCFA_BACSUBacillus subtilis lcfA31.159.9534long-chain-fatty-acid—CoA ligase
3183818302036301512525gp: SCJ4_28Streptomyces coelicolor A3(2)33.965.4127transcriptional regulator
SCJ4.28c
3193819302167303099933sp: FABG_BACSUBacillus subtilis fabG41.072.52513-oxoacyl-(acyl-carrier-protein)
reductase
3203820303133304074942sp: FLUG_EMENIEmericella nidulans fluG27.252.0254glutamine synthetase
32138213040703052631194prf: 2512386AArabidopsis thaliana atg638.866.5394short-chain acyl CoA oxidase
3223822305288305758471sp: NODN_RHILVRhizobium leguminosarum nodN45.872.6153nodulation protein
3233823305858306700843pir: F70790Mycobacterium tuberculosis41.272.4272hydrolase
H37Rv Rv3677c
32438243063673051951173
3253825306800307504705
3263826307462306782681prf: 2323349AVibrio cholerae crp30.965.7207cAMP receptor protein
3273827307918307727192
3283828307955308734780sp: UVEN_MICLUMicrococcus luteus pdg57.577.1240ultraviolet N-glycosylase/AP lyase
3293829308745309302558pir: B70790Mycobacterium tuberculosis34.658.3211cytochrome c biogenesis protein
H37Rv Rv3673c
3303830309370310038669sp: YEAB_ECOLIEscherichia coli K12 yeaB30.756.3192hypothetical protein
33138313101353113251191pir: H70789Mycobacterium tuberculosis38.671.0396serine proteinase
H37Rv Rv3671c
3323832312891311899993prf: 2411250ACorynebacterium sp. C12 cEH29.652.1280epoxide hydrolase
3333833313457312909549pir: F70789Mycobacterium tuberculosis46.877.6156hypothetical membrane protein
H37Rv Rv3669
3343834314590313625966pir: S72914Mycobacterium leprae29.665.5287phosphoserine phosphatase
MTCY20G9.32C. serB
33538353149803160021023pir: E70788Mycobacterium tuberculosis35.060.2349hypothetical protein
H37Rv Rv3660c
33638363161103171321023pir: C44020Escherichia coli trbB32.966.5319conjugal transfer region protein
3373837316964316350615
3383838317078317893816pir: C70788Mycobacterium tuberculosis30.563.7262hypothetical membrane protein
H37Rv Rv3658c
3393839317920318465546pir: B70788Mycobacterium tuberculosis33.864.2201hypothetical protein
H37Rv Rv3657c
3403840318492318689198pir: A70788Mycobacterium tuberculosis47.584.859hypothetical protein
H37Rv Rv3656c
3413841318696319013318
3423842318958318545414
3433843318991319335345
34438443216903193362355sp: YPRA_BACSUBacillus subtilis yprA33.866.1764ATP-dependent RNA helicase
3453845322007322207201sp: CSP_ARTGOArthrobacter globiformis SI5568.788.167cold shock protein
csp
3463846322216321992225
34738473229103258972988pir: G70563Mycobacterium tuberculosis61.781.6977DNA topoisomerase I
H37Rv Rv3646c topA
3483848325904326614711
34938493277353266951041sp: CYAB_STIAUStigmatella aurantiaca B17R2032.762.4263adenylate cyclase
cyaB
35038503282833295391257sp: DP3X_BACSUBacillus subtilis dnaX25.352.7423DNA polymerase III subunit
tau/gamma
3513851329748329909162
3523852329933330376444gp: AE002103_3Ureaplasma urealyticum uu03332.659.0144hypothetical protein
3533853330973331533561gp: AE001882_8Deinococcus radiodurans39.063.4172hypothetical protein
DR0202
3543854331552332433882sp: RLUC_ECOLIEscherichia coli K12 rluC43.665.0314ribosomal large subunit
pseudouridine synthase C
35538553329193345621644sp: BGLX_ERWCHErwinia chrysanthemi D1 bgxA34.860.2558beta-glucosidase/xylosidase
35638563329653349531989gp: AF090429_2Azospirillum irakense salB38.661.4101beta-glucosidase
35738573350093361121104sp: FADH_AMYMEAmycolatopsis methanolica66.686.5362NAD/mycothiol-dependent
formaldehyde dehydrogenase
3583858335805335185621
3593859336212336748537sp: YTH5_RHOSNRhodococcus erythropolis orf532.547.5160metallo-beta-lactamase superfamily
3603860336781337449669sp: FABG_ECOLIEscherichia coli K12 fabG25.955.82513-oxoacyl-(acyl-carrier-protein)
reductase
36138613375393387681230gp: AF148322_1Streptomyces viridifaciens vlmF26.356.4415valanimycin resistant protein
3623862338793339725933prf: 2512357BActinoplanes sp. acbB33.866.3320dTDP-glucose 4,6-dehydratase
3633863340569340195375pir: A70562Mycobacterium tuberculosis59.388.9108hypothetical protein
H37Rv Rv3632
3643864341327340569759sp: YC22_METJAMethanococcus jannaschii JAL-33.966.5230dolichol phosphate mannose
1 MJ1222synthase
36538653413473423751029
36638663424173434511035sp: YEFJ_ECOLIEscherichia coli K12 yefJ25.857.3260nucleotide sugar synthetase
36738673436363457172082sp: USHA_SALTYSalmonella typhimurium ushA26.154.4586UDP-sugar hydrolase
3683868345975345814162
3693869346460346110351
37038703480193469611059sp: ADH_MYCTUMycobacterium tuberculosis52.274.9343NADP-dependent alcohol
H37Rv adhCdehydrogenase
3713871348952348098855sp: RFBA_SALANSalmonella anatum M32 rfbA62.884.9285glucose-1-phosphate
thymidylyltransferase
37238723503103489521359gp: D78182_5Streptococcus mutans rmlC49.574.0192dTDP-4-keto-L-rhamnose reductase
37338733514433503131131sp: RMLB_STRMUStreptococcus mutans XC rmlB61.883.4343dTDP-glucose 4,6-dehydratase
3743874351948351370579sp: NOX_THETHThermus aquaticus HB8 nox35.461.2206NADH dehydrogenase
3753875352693353637945prf: 2510361AStaphylococcus aureus sirA33.266.5325Fe-regulated protein
3763876354387353749639
37738773559063545991308sp: Y17M_MYCTUMycobacterium tuberculosis37.468.3423hypothetical membrane protein
H37Rv Rv3630
37838783572283558491380gp: SC5F2A_19Streptomyces coelicolor34.162.5461metallopeptidase
SC5F2A.19c
37938793593543572372118prf: 2502226ASphingomonas capsulata28.456.4708prolyl endopeptidase
3803880360334359762573
38138813619053608141092gp: SCF43_2Streptomyces coelicolor A3(2)26.046.0258hypothetical membrane protein
38238823631513620571095gsp: W56155Corynebacterium50.776.6363cell surface layer protein
ammoniagenes ATCC 6872
38338833638243652571434prf: 2404346BAcinetobacter johnsonii ptk28.557.2453autophosphorylating protein Tyr
kinase
3843884365250365852603prf: 2404346AAcinetobacter johnsonii ptp39.268.6102protein phosphatase
3853885365855366838984
38638863668323686431812sp: CAPD_STAAUStaphylococcus aureus M capD33.065.7613capsular polysaccharide
biosynthesis
3873887368642367701942PRF: 2109288XVibrio cholerae41.051.090ORF 3
38838883686473698011155prf: 2423410LCampylobacter jejuni wlaK37.168.3394lipopolysaccharide biosynthesis/
aminotransferase
3893889369794370405612gp: AF014804_1Neisseria meningitidis pglB54.675.0196pilin glycosylation protein
39038903706133717731161sp: CAPM_STAAUStaphylococcus aureus M capM33.469.2380capsular polysaccharide
biosynthesis
39138913719293734191491pir: S67859Xanthomonas campestris gumJ34.369.8504lipopolysaccharide biosynthesis/
export protein
39238923735003748131314sp: MURA_ENTCLEnterobacter cloacae murA31.464.6427UDP-N-acetylglucosamine 1-
carboxyvinyltransferase
39338933748333758371005sp: MURB_BACSUBacillus subtilis murB34.868.5273UDP-N-
acetylenolpyruvoylglucosamine
reductase
39438943758423768761035gp: VCLPSS_9Vibrio cholerae ORF39 × 232.057.3356sugar transferase
3953895377683377832150prf: 2211295ACorynebacterium glutamicum60.479.353transposase
3963896378093378227135
3973897378185378511327pir: S43613Corynebacterium glutamicum75.794.370transposase (insertion sequence
ATCC 31831IS31831)
3983898378562378287276
39938993798373786681170pir: G70539Mycobacterium tuberculosis28.057.4404hypothetical protein
H37Rv Rv1565c
4003900380842379850993gsp: W37352Pseudomonas aeruginosa PAO134.560.2354acetyltransferase
psbC
4013901381265381495231PIR: S60890Corynebacterium glutamicum44.053.065hypothetical protein B
40239023819483831081161sp: UDG8_ECOLIEscherichia coli ugd63.789.7388UDP-glucose 6-dehydrogenase
4033903383768383496273
40439043851903839821209
4053905386195385374822gp: AF172324_3Escherichia coli wbnA32.165.0243glycosyl transferase
4063906386556387200645gp: AB008676_13Escherichia coli 0157 wbhH33.062.0221acetyltransferase
4073907387657387463195
40839083876923890981407gp: CGLPD_1Corynebacterium glutamicum99.6100.0469dihydrolipoamide dehydrogenase
ATCC 13032 lpd
4093909389248390168921pir: JC4985Xanthomonas campestris41.768.1295UTP—glucose-1-phosphate
uridylyltransferase
4103910390233390730498gp: PAU49666_2Pseudomonas aeruginosa PAO143.871.9153regulatory protein
orfX
41139113922083907871422pir: E70828Mycobacterium tuberculosis57.081.3477transcriptional regulator
H37Rv Rv0465c
4123912392705393475771gp: SCM10_12Streptomyces coelicolor A3(2)34.867.4230cytochrome b subunit
SCM10.12c
41339133936393955131875pir: A27763Bacillus subtilis sdhA32.461.2608succinate dehydrogenase
flavoprotein
4143914395426396262837gp: BMSDHCAB_4Paenibacillus macerans sdhB27.556.2258succinate dehydrogenase subunit B
4153915396315396650336
4163916396672396932261
4173917397040396411630
418391839773039782596
4193919397884398222339
4203920398206397232975gp: SCC78_5Streptomyces coelicolor26.349.8259hypothetical protein
SCC78.05
42139213983293995791251sp: YJIN_ECOLIEscherichia coli K12 yjiN32.764.3431hypothetical protein
4223922399598400017420
4233923400039400341303
4243924400473401150678sp: TCMR_STRGAStreptomyces glaucescens26.453.8197tetracenomycin C transcription
GLA.0 tcmRrepressor
4253925401050401253204
42639264011504027961647gp: AF164961_8Streptomyces fradiae T#271736.174.6499transporter
urdJ
42739274027994044301632gp: AF164961_8Streptomyces fradiae T#271739.674.6508transporter
urdJ
4283928405419404508912sp: PURU_CORSPCorynebacterium sp. P-1 purU40.972.7286formyltetrahydrofolate deformylase
4293929405480406145666sp: DEOC_BACSUBacillus subtilis deoC38.574.0208deoxyribose-phosphate aldolase
4303930406310406161150
4313931406417405521897
4323932406550407416867prf: 2413441KMycobacterium avium GIR1026.853.6280hypothetical protein
mav346
4333933407708407409300pir: A70907Mycobacterium tuberculosis58.785.992hypothetical protein
H37Rv Rv0190
4343934408546409145600
43539354099754077112265sp: CTPB_MYCLEMycobacterium leprae ctpB45.775.3748cation-transporting P-type ATPase B
4363936410476410027450
43739374106834125451863sp: AMYH_YEASTSaccharomyces cerevisiae27.356.1626glucan 1,4-alpha-glucosidase
S288C YIR019C sta1
43839384125574136331077gp: AF109162_1Corynebacterium diphtheriae57.283.6348hemin-binding periplasmic protein
hmuT
43939394136434147101068gp: AF109162_2Corynebacterium diphtheriae65.290.3330ABC transporter
hmuU
4403940414714415526813gp: AF109162_3Corynebacterium diphtheriae63.885.0254ABC transporter ATP-binding protein
hmuV
4413941415643416599957gp: SCC75A_17Streptomyces coelicolor C75A28.656.4266hypothetical protein
SCC75A.17c
4423942416603417439837gp: SCC75A_17Streptomyces coelicolor C75A32.661.6258hypothetical protein
SCC75A.17c
4433943418354417545810
4443944419253418441813
4453945419757419257501
44639464197854208851101gp: ECOMURBA_1Escherichia coli RDD012 murB30.158.4356UDP-N-acetylpyruvoylglucosamine
reductase
4473947420866421516651
4483948421043420309735
4493949421858422031174
45039504237934220901704sp: LCFA_BACSUBacillus subtilis lcfA35.568.1558long-chain-fatty-acid—CoA ligase
45139514238784251311254gp: SC2G5_6Streptomyces coelicolor33.958.7416transferase
SC2G5.06
4523952425177425920744sp: PMGY_STRCOStreptomyces coelicolor A3(2)70.784.2246phosphoglycerate mutase
gpm
45339534259344271721239prf: 2404434AMycobacterium bovis senX349.274.8417two-component system sensor
histidine kinase
4543954427172427867696prf: 2404434BMycobacterium bovis BCG75.890.9231two-component response regulator
regX3
4553955428561429439879
45639564320234294382586gp: SCE25_30Streptomyces coelicolor A3(2)31.360.7921ABC transporter ATP-binding protein
SCE25.30
4573957433028432126903sp: YV21_MYCTUMycobacterium tuberculosis45.066.9269cytochrome P450
H37Rv RV3121
4583958433062433988927prf: 2512277APseudomonas aeruginosa ppx28.857.8306exopolyphosphatase
4593959434010434822813sp: YV23_MYCTUMycobacterium tuberculosis28.857.3302hypothetical membrane protein
H37Rv Rv0497
4603960434886435695810sp: PROC_CORGLCorynebacterium glutamicum100.0100.0269pyrroline-5-carboxylate reductase
ATCC 17965 proC
46139614349864338651122gp: D88733_1Equine herpesvirus 1 ORF7125.452.0394membrane glycoprotein
4623962435940436137198pir: S72921Mycobacterium leprae76.494.655hypothetical protein
B2168_C1_172
4633963436321436103219
464396443646343656199gp: SCE68_25Streptomyces coelicolor89.7100.029hypothetical protein
SCE68.25c
4653965436573436764192
4663966437233437850618
46739674380444369801065pir: S72914Mycobacterium leprae51.077.4296phosphoserine phosphatase
MTCY20G9.32C. serB
4683968438179438424246sp: YV35_MYCTUMycobacterium tuberculosis40.566.274hypothetical protein
H37Rv Rv0508
4693969438294438037258
47039704385164399041389sp: HEM1_MYCLEMycobacterium leprae hemA44.474.3455glutamyl-tRNA reductase
4713971439909440814906pir: S72887Mycobacterium leprae hem3b50.775.3308hydroxymethylbilane synthase
4723972441220441591372
4733973442482441601882sp: CATM_ACICAAcinetobacter calcoaceticus27.157.6321cat operon transcriptional regulator
catM
47439744427584441581401sp: SHIA_ECOLIEscherichia coli K12 shiA35.572.2417shikimate transport protein
47539754441854460381854sp: 3SHD_NEUCRNeurospora crassa qa428.257.93093-dehydroshikimate dehydratase
4763976446538447386849gp: AF124518_2Corynebacterium glutamicum98.298.6282shikimate dehydrogenase
ASO19 aroE
4773977447670447398273
47839784491794481301050sp: POTG_ECOLIEscherichia coli K12 potG34.768.6363putrescine transport protein
4793979449714449100615
48039804508264491831644sp: SFUB_SERMASerratia marcescens sfuB25.155.2578iron(III)-transport system permease
protein
48139814508494519611113
48239824518954508371059gp: SHU75349_1Brachyspira hyodysenteriae bitA25.159.9347periplasmic-iron-binding protein
48339834526614544301770pir: S72909Mycobacterium leprae cysG46.571.6486uroporphyrin-III C-methyltransferase
4843984454450454875426
48539854549674559831017sp: HEM2_STRCOStreptomyces coelicolor A3(2)60.883.1337delta-aminolevulinic acid
hemBdehydratase
4863986456016456597582
4873987456641457150510
48839884573574599002544sp: CTPB_MYCLEMycobacterium leprae ctpB27.456.5858cation-transporting P-type ATPase B
4893989459425458583843
49039904600204610931074sp: DCUP_STRCOStreptomyces coelicolor A3(2)55.076.7364uroporphyrinogen decarboxylase
hemE
49139914611124624551344sp: PPOX_BACSUBacillus subtilis hemY28.059.9464protoporphyrinogen IX oxidase
49239924625574638671311sp: GSA_MYCLEMycobacterium leprae hemL61.783.5425glutamate-1-semialdehyde 2,1-
aminomutase
4933993463867464472606sp: PMG2_ECOLIEscherichia coli K12 gpmB28.062.7161phosphoglycerate mutase
4943994464482465102621pir: A70545Mycobacterium tuberculosis44.771.2208hypothetical protein
H37Rv Rv0526
4953995465118465909792pir: B70545Mycobacterium tuberculosis53.585.3245cytochrome c-type biogenesis
H37Rv ccsAprotein
49639964659494675711623pir: C70545Mycobacterium tuberculosis50.776.0533hypothetical membrane protein
H37Rv Rv0528
49739974676484686581011pir: D70545Mycobacterium tuberculosis44.177.8338cytochrome c biogenesis protein
H37Rv ccsB
4983998469370470170801
4993999470184470654471pir: G70790Mycobacterium tuberculosis38.969.4144transcriptional regulator
H37Rv Rv3678c pb5
5004000471013470657357prf: 2420312AStaphylococcus aureus zntR31.172.290Zn/Co transport repressor
5014001471420471121300
5024002471515471847333pir: F70545Mycobacterium tuberculosis39.078.182hypothetical membrane protein
H37Rv Rv0531
5034003472808471915894sp: MENA_ECOLIEscherichia coli K12 menA33.661.53011,4-dihydroxy-2-naphthoate
octaprenyltransferase
5044004472948473811864gp: AF125164_6Bacteroides fragilis wcgB32.462.6238glycosyl transferase
50540054751364738141323prf: 2423270BRhizobium trifolii matB25.451.5421malonyl-CoA-decarboxylase
5064006475407474997411sp: YQJF_ECOLIEscherichia coli K12 yqjF35.365.5139hypothetical membrane protein
50740074770484754891560pir: S27612Pseudomonas putida 50.476.0520ketoglutarate semialdehyde
dehydrogenase
5084008477995477048948sp: KDGD_PSEPUPseudomonas putida KDGDH48.575.63035-dehydro-4-deoxyglucarate
dehydratase
5094009478970478092879sp: ALSR_BACSUBacillus subtilis 168 alsR36.966.2293als operon regulatory protein
5104010479303478989315pir: B70547Mycobacterium tuberculosis33.064.994hypothetical protein
H37Rv Rv0543c
5114011480154480597444
5124012480201479452750gp: SSP277295_9Sphingomonas sp. LB126 fldB28.154.72672-pyrone-4,6-dicarboxylic acid
5134013480624480208417
5144014481001480624378
5154015481391481131261
51640164826684813941275pir: D70547Mycobacterium tuberculosis60.083.2410low-affinity inorganic phosphate
H37Rv pitAtransporter
5174017483587483366222
5184018483942483637306
5194019485062484106957sp: MENB_BACSUBacillus subtilis menB48.570.3293naphthoate synthase
5204020485384485986603gp: AE001957_12Deinococcus radiodurans57.982.7202peptidase E
DR1070
5214021485385485077309pir: C70304Aquifex aeolicus VF5 phhB37.768.877pterin-4a-carbinolamine dehydratase
52240224860014870141014pir: D70548Mycobacterium tuberculosis54.076.7335muconate cycloisomerase
H37Rv Rv0553 menC
52340234870284886561629sp: MEND_BACSUBacillus subtilis menD29.454.06062-oxoglutarate decarboxylase and 2-
succinyl-6-hydroxy-2,4-
cyclohexadiene-1-carboxylate
synthase
5244024488660489100441pir: G70548Mycobacterium tuberculosis37.264.9148hypothetical membrane protein
H37Rv Rv0556
52540254892094904471239pir: H70548Mycobacterium tuberculosis22.854.2408alpha-D-mannose-alpha(1-6)
H37Rv pimBphosphatidyl myo-inositol
monomannoside transferase
52640264905804919381359sp: CYCA_ECOLIEscherichia coli K12 cycA66.289.9447D-serine/D-alanine/glycine
transporter
5274027491966492655690sp: UBIE_ECOLIEscherichia coli K12 ubiE37.166.7237ubiquinone/menaquinone
biosynthesis methyltransferase
5284028492915493583669
52940294939164926451272pir: D70549Mycobacterium tuberculosis49.076.7412oxidoreductase
H37Rv Rv0561c
53040304940614951101050sp: HEP2_BACSTBacillus stearothermophilus39.267.1316heptaprenyl diphosphate synthase
ATCC 10149 hepTcomponent II
5314031496810497142333gp: AF130462_2Corynebacterium glutamicum100.0100.0111preprotein translocase SecE subunit
ATCC 13032 secE
5324032497374498327954gp: AF130462_3Corynebacterium glutamicum100.0100.0318transcriptional antiterminator protein
ATCC 13032 nusG
5334033498598499032435gp: AF130462_4Corynebacterium glutamicum100.0100.014550S ribosomal protein L11
ATCC 13032 rplK
5344034499162499869708gp: AF130462_5Corynebacterium glutamicum100.0100.023650S ribosomal protein L1
ATCC 13032 rplA
53540355014364999251512gp: SC5H4_2Streptomyces coelicolor23.150.2564regulatory protein
SC5H4.02
53640365015775029201344sp: GABT_MYCTUMycobacterium tuberculosis60.582.44434-aminobutyrate aminotransferase
H37Rv RV2589 gabT
53740375029255042831359sp: GABD_ECOLIEscherichia coli K12 gabD40.871.8461succinate-semialdehyde
dehydrogenase (NAD(P)+)
5384038503739503272468GP: ABCARRA_2Azospirillum brasilense carR32.038.0150novel two-component regulatory
system
53940395043795055691191sp: TYRP_ECOLIEscherichia coli K12 o341#725.549.9447tyrosine-specific transport protein
tyrP
54040405056985076471950sp: CTPG_MYCTUMycobacterium tuberculosis33.264.4615cation-transporting ATPase G
H37Rv RV1992C ctpG
54140415076695090811413sp: P49_STRLIStreptomyces lividans P4940.266.2468hypothetical protein or
dehydrogenase
5424042509094509696603
5434043509998510510513sp: RL10_STRGRStreptomyces griseus N2-3-1152.984.717050S ribosomal protein L10
rplJ
5444044510591510974384sp: RL7_MYCTUMycobacterium tuberculosis72.389.213050S ribosomal protein L7/L12
H37Rv RV0652 rplL
5454045511126510989138
5464046511536512507972pir: A70962Mycobacterium tuberculosis25.855.5283hypothetical membrane protein
H37Rv Rv0227c
54740475129135164073495sp: RPOB_MYCTUMycobacterium tuberculosis75.490.41180DNA-directed RNA polymerase beta
H37Rv RV0667 rpoBchain
54840485164945204923999sp: RPOC_MYCTUMycobacterium tuberculosis72.988.71332DNA-directed RNA polymerase beta
H37Rv RV0668 rpoCchain
5494049519277518696582GP: AF121004_1Mycobacterium tuberculosis39.052.0169hypothetical protein
H37Rv Jv0166c
5504050520671520850180
5514051520865521644780gp: SCJ9A_15Streptomyces coelicolor A3(2)39.263.8232DNA-binding protein
SCJ9A.15c
5524052522476521679798sp: YT08_MYCTUMycobacterium tuberculosis29.357.7215hypothetical protein
H37Rv RV2908C
5534053522694523059366sp: RS12_MYCITMycobacterium intracellulare90.997.512130S ribosomal protein S12
rpsL
5544054523069523533465sp: RS7_MYCSMMycobacterium smegmatis81.894.815430S ribosomal protein S7
LR222 rpsG
55540555238965260102115sp: EFG_MICLUMicrococcus luteus fusA71.788.9709elongation factor G
55640565260705239112160
5574057526156526013144
5584058527121526894228GSP: Y37841Chlamydia trachomatis56.078.044lipoprotein
5594059527759527607153
5604060528040528768729
5614061529570528779792sp: FEPC_ECOLIEscherichia coli K12 fepC56.283.7258ferric enterobactin transport ATP-
binding protein
56240625306265295921035sp: FEPG_ECOLIEscherichia coli K12 fepG45.677.8329ferric enterobactin transport protein
56340635317825307481035sp: FEPD_ECOLIEscherichia coli K12 fepD48.180.6335ferric enterobactin transport protein
5644064532008532523516gp: CTACTAGEN_1Thermoanaerobacterium56.679.3145butyryl-CoA: acetate coenzyme A
thermosaccharolyticum actAtransferase
5654065533099533401303sp: RS10_PLAROPlanobispora rosea ATCC84.299.010130S ribosomal protein S10
53733 rpsJ
5664066533437534090654sp: RL3_MYCBOMycobacterium bovis BCG rplC66.589.621250S ribosomal protein L3
5674067534087533401687
5684068534090534743654sp: RL4_MYCBOMycobacterium bovis BCG rplD71.290.121250S ribosomal protein L4
5694069534746535048303sp: RL23_MYCBOMycobacterium bovis BCG rplW74.090.69650S ribosomal protein L23
5704070535072534746327
5714071535076535915840sp: RL2_MYCLEMycobacterium bovis BCG rplB80.792.928050S ribosomal protein L2
5724072535935536210276sp: RS19_MYCTUMycobacterium tuberculosis87.098.99230S ribosomal protein S19
H37Rv Rv0705 rpsS
5734073536183535899285
5744074536217536576360sp: RL22_MYCTUMycobacterium tuberculosis74.391.710950S ribosomal protein L22
H37Rv Rv0706 rplV
5754075536579537322744sp: RS3_MYCBOMycobacterium bovis BCG rpsC77.491.223930S ribosomal protein S3
5764076537328537741414sp: RL16_MYCBOMycobacterium bovis BCG rplP69.388.313750S ribosomal protein L16
5774077537744537971228sp: RL29_MYCBOMycobacterium bovis BCG rpmC65.788.16750S ribosomal protein L29
5784078537977538252276sp: RS17_MYCBOMycobacterium bovis BCG rpsQ69.589.08230S ribosomal protein S17
5794079538267537974294
5804080538698538381318
5814081539413538718696
5824082539741540106366sp: RL14_MYCTUMycobacterium tuberculosis83.695.112250S ribosomal protein L14
H37Rv Rv0714 rplN
5834083540112540423312sp: RL24_MYCTUMycobacterium tuberculosis76.291.410550S ribosomal protein L24
H37Rv Rv0715 rplX
5844084540426540998573sp: RL5_MICLUMicrococcus luteus rplE73.692.318350S ribosomal protein L5
58540855410485420791032
5864086542896542090807sp: 2DKG_CORSPCorynebacterium sp.52.374.22602,5-diketo-D-gluconic acid reductase
5874087543412542921492
5884088544329543415915sp: FDHD_WOLSUWolinella succinogenes fdhD28.959.7298formate dehydrogenase chain D
5894089544670544335336gp: SCGD3_29Streptomyces coelicolor A3(2)37.268.194molybdopterin-guanine dinucleotide
SCGD3.29cbiosynthesis protein
59040905468895447572133sp: FDHF_ECOLIEscherichia coli fdfF24.353.4756formate dehydrogenase H or alpha
chain
5914091547329548084756
5924092548990548187804
59340935506515489901662sp: YC81_MYCTUMycobacterium tuberculosis26.952.6624ABC transporter ATP-binding protein
H37Rv Rv1281c oppD
59440945518445506991146
59540955529275518541074
59640965541295529481182pir: E69424Archaeoglobus fulgidus AF139824.750.4405hypothetical protein
5974097554919554452468gp: AE001931_13Deinococcus radiodurans42.766.7150hypothetical protein
DR0763
5984098555331555726396pir: S29885Micrococcus luteus 75.897.713230S ribosomal protein S8
5994099555749556282534pir: S29886Micrococcus luteus 59.287.717950S ribosomal protein L6
6004100556289556690402sp: RL18_MICLUMicrococcus luteus rplR67.390.911050S ribosomal protein L18
6014101556734557366633sp: RS5_MICLUMicrococcus luteus rpsE67.888.317130S ribosomal protein S5
6024102557373557555183sp: RL30_ECOLIEscherichia coli K12 rpmJ54.676.45550S ribosomal protein L30
6034103557565558008444sp: RL15_MICLUMicrococcus luteus rplO66.487.414350S ribosomal protein L15
6044104557588556860729
6054105558517558197321prf: 2204281AStreptomyces coelicolor msdA46.968.8128methylmalonic acid semialdehyde
dehydrogenase
6064106558969558607363
6074107559805560260456GP: ABCARRA_2Azospirillum brasilense carR47.052.0125novel two-component regulatory
system
60841085606345591441491prf: 2516398ERhodococcus rhodochrous41.771.5487aldehyde dehydrogenase or betaine
plasmid pRTL1 orf5aldehyde dehydrogenase
6094109561368560634735
6104110562632562937306
61141115626335613681266prf: 2411257BSphingomonas sp. redA241.171.6409reductase
6124112562963562646318prf: 2313248BRhodobacter capsulatus fdxE47.766.41072Fe2S ferredoxin
6134113563736562993744gp: PPU24215_2Pseudomonas putida cymB35.870.8257p-cumic alcohol dehydrogenase
6144114563871564083213PIR: H72754Aeropyrum pernix K1 APE002950.056.050hypothetical protein
61541155654715637321740pir: JC4176Pyrococcus furiosus Vc1 DSM22.945.0629phosphoenolpyruvate synthetase
3638 ppsA
61641165667595656801080pir: JC4176Pyrococcus furiosus Vc1 DSM38.666.7378phosphoenolpyruvate synthetase
3638 ppsA
61741175680885667991290prf: 2104333GRhodococcus erythropolis thcB34.865.2422cytochrome P450
6184118569075568272804prf: 2512309AErwinia carotovora carotovora28.566.0256transcriptional repressor
kdgR
6194119570774571316543sp: KAD_MICLUMicrococcus luteus adk48.981.0184adenylate kinase
6204120571367570756612
6214121571476572267792sp: AMPM_BACSUBacillus subtilis 168 map43.174.7253methionine aminopeptidase
6224122572349573176828
6234123573407573622216pir: F69644Bacillus subtilis infA77.086.072translation initiation factor IF-1
6244124573816574181366prf: 2505353BThermus thermophilus HB866.491.012230S ribosomal protein S13
rps13
6254125574187574588402sp: RS11_STRCOStreptomyces coelicolor A3(2)81.393.313430S ribosomal protein S11
SC6G4.06. rpsK
6264126574615575217603prf: 2211287FMycobacterium tuberculosis82.693.913230S ribosomal protein S4
H37Rv RV3458C rpsD
62741275753385763511014sp: RPOA_BACSUBacillus subtilis 168 rpoA51.177.8311RNA polymerase alpha subunit
6284128575366575211156
6294129576410576898489sp: RL17_ECOLIEscherichia coli K12 rplQ51.677.112250S ribosomal protein L17
6304130577057577923867sp: TRUA_ECOLIEscherichia coli k12 truA37.061.1265pseudouridylate synthase A
63141315780335804292397pir: G70695Mycobacterium tuberculosis24.851.2786hypothetical membrane protein
H37Rv Rv3779
6324132580891580436456
6334133581221580919303
63441345814065826621257pir: A70836Mycobacterium tuberculosis27.453.8485hypothetical protein
H37Rv Rv0283
63541355826845842281545sp: DIM_ARATHArabidopsis thaliana CV DIM22.850.9505cell elongation protein
63641365842685856201353sp: CFA_ECOLIEscherichia coli K12 cfa30.756.0423cyclopropane-fatty-acyl-phospholipid
synthase
6374137585823586248426gp: SCL2_30Streptomyces coelicolor A3(2)28.059.0100hypothetical membrane protein
SCL2.30c
63841385877575863991359sp: ELYA_BACAOBacillus alcalophilus31.358.0273high-alkaline serine proteinase
63941395890155876451371pir: T10930Streptomyces coelicolor A3(2)24.050.6516hypothetical membrane protein
SC3C3.21
64041405892965928623567pir: E70977Mycobacterium tuberculosis65.038.41260hypothetical membrane protein
H37Rv Rv3447c
6414141590411589590822
6424142590560589898663
6434143592862593761900
6444144593935594258324pir: C70977Mycobacterium tuberculosis31.169.9103hypothetical protein
H37Rv Rv3445c
6454145594293594580288prf: 2111376AMycobacterium tuberculosis36.381.380early secretory antigen target ESAT-
6 protein
6464146594939595379441sp: RL13_STRCOStreptomyces coelicolor A3(2)58.682.114550S ribosomal protein L13
SC6G4.12. rplM
6474147595382595927546sp: RS9_STRCOStreptomyces coelicolor A3(2)49.272.418130S ribosomal protein S9
SG6G4.13. rpsl
64841485961095974491341prf: 2320260AStaphylococcus aureus 48.976.4450phosphoglucosamine mutase
femR315
6494149597892598194303
65041505981945997021509pir: S75138Synechocystis sp. PCC680329.345.6318hypothetical protein
slr1753
6514151599350598778573
6524152599699599932234
6534153600876600022855pir: S73000Mycobacterium leprae44.072.2259hypothetical protein
B229_F1_20
65441546009716020531083sp: ALR_MYCTUMycobacterium tuberculosis41.668.5368alanine racemase
H37Rv RV3423C alr
6554155602080602574495sp: Y097_MYCTUMycobacterium tuberculosis48.778.6154hypothetical protein
H37Rv Rv3422c
65641566028116044091599sp: YIDE_ECOLIEscherichia coli K12 yidE28.966.2550hypothetical membrane protein
65741576044706057081239gp: PSJ00161_1Propionibacterium shermanii pip51.377.6411proline iminopeptidase
6584158605718606392675sp: Y098_MYCTUMycobacterium tuberculosis52.275.4207hypothetical protein
H37Rv Rv3421c
6594159606392606898507sp: RIMI_ECOLIEscherichia coli K12 riml30.359.9132ribosomal-protein-alanine N-
acetyltransferase
66041606069056079361032sp: GCP_PASHAPasteurella haemolytica46.175.2319O-sialoglycoprotein endopeptidase
SEROTYPE A1 gcp
66141616079586096791722sp: Y115_MYCTUMycobacterium tuberculosis38.459.4571hypothetical protein
H37Rv Rv3433c
6624162609747610175429
6634163610268609816453
6644164610348610644297sp: CH10_MYCTUMycobacterium tuberculosis76.094.0100heat shock protein groES
H37Rv RV3418C mopB
66541656106596122721614sp: CH61_MYCLEMycobacterium leprae63.385.1537heat shock protein groEL
B229_C3_248 groE1
6664166611200610946255GP: MSGTCWPA_1Mycobacterium tuberculosis50.056.076hypothetical protein
66741676122666111091158GP: MSGTCWPA_3Mycobacterium tuberculosis34.045.0138hypothetical protein
6684168612714612418297gp: AF073300_1Mycobacterium smegmatis64.988.394regulatory protein
whiB3
6694169613156613719564sp: Y09F_MYCTUMycobacterium tuberculosis55.281.6174RNA polymerase sigma factor
H37Rv Rv3414c sigD
67041706137226147471026
6714171615180614803378sp: Y09H_MYCLEMycobacterium leprae41.469.8116hypothetical protein
B1620_F3_131
67241726153366168531518gp: AB003154_1Corynebacterium80.893.9504IMP dehydrogenase
ammoniagenes ATCC 6872
guaB
6734173616231615605627PIR: F71456Pyrococcus horikoshii PH030839.053.0146hypothetical protein
67441746169736180941122gp: AB003154_2Corynebacterium70.986.1381IMP dehydrogenase
ammoniagenes ATCC 6872
6754175619013618093921sp: YBIF_ECOLIEscherichia coli K12 ybiF38.067.5274hypothetical membrane protein
6764176619086619994909prf: 1516239ABacillus subtilis gltC29.058.4262glutamate synthetase positive
regulator
67741776200046215721569sp: GUAA_CORAMCorynebacterium81.692.8517GMP synthetase
ammoniagenes guaA
6784178620926620264663
6794179621717622157441
6804180622269622457189
68141816236356224601176gp: SCD63_22Streptomyces coelicolor A3(2)20.539.6513hypothetical membrane protein
68241826238006249391140gp: SC6E10_15Streptomyces coelicolor A3(2)26.848.7411two-component system sensor
SC6E10.15chistidine kinase
6834183624985625674690sp: DEGU_BACSUBacillus subtilis 168 degU33.565.1218transcriptional regulator or
extracellular proteinase response
regulator
6844184625677626000324
6854185626558626070489
6864186627539626577963
6874187627727628551825pir: B70975Mycobacterium tuberculosis30.964.2201hypothetical protein
H37Rv Rv3395c
68841886285516301401590pir: A70975Mycobacterium tuberculosis37.564.1563hypothetical protein
H37Rv Rv3394c
6894189630810630151660
6904190630949631809861gp: SC5B8_20Streptomyces coelicolor A3(2)33.862.9275hypothetical protein
SC5B8.20c
6914191632684631824861gp: AE001935_7Deinococcus radiodurans27.858.3288hypothetical membrane protein
DR0809
6924192633079632690390
6934193633474633079396gp: MMU92075_3Mycobacterium marinum36.867.495hypothetical membrane protein
69441946351756335321644gp: AF139916_3Brevibacterium linens ATCC50.476.2524phytoene desaturase
9175 crtl
6954195636089635178912gp: AF139916_2Brevibacterium linens ATCC42.071.2288phytoene synthase
9175 crtB
69641966382786360892190gp: SCF43A_29Streptomyces coelicolor A3(2)48.675.6722transmembrane transport protein
SCF43A.29c
69741976394626383171146gp: AF139916_11Brevibacterium linens crtE32.763.8367geranylgeranyl pyrophosphate
(GGPP) synthase
6984198639624640208585gp: AF139916_14Brevibacterium linens 38.368.1188transcriptional regulator (MarR
family)
6994199640879640232648sp: BLC_CITFRCitrobacter freundii blc OS60 blc33.162.1145outer membrane lipoprotein
70042006411336425571425gp: AF139916_1Brevibacterium linens 48.774.2462hypothetical protein
70142016439596425561404gp: AF139916_5Brevibacterium linens ATCC40.063.2497DNA photolyase
9175 cpd1
7024202644026644778753gp: AF155804_7Streptococcus suis cps1K25.953.7205glycosyl transferase
70342036475906451762415gp: SCE25_30Streptomyces coelicolor A3(2)24.354.9897ABC transporter
SCE25.30
7044204648309647593717prf: 2420410PBacillus subtilis 168 yvrO35.472.2223ABC transporter
7054205648467648315153
7064206649105648440666prf: 2320284DHelicobacter pylori abcD35.975.2206ABC transporter
7074207649342650187846
70842086501936491141080sp: ABC_ECOLIEscherichia coli TAP90 abc43.675.4346ABC transporter
7094209651288650392897sp: HLPA_HAEINHaemophilus influenzae 28.767.2268lipoprotein
SEROTYPE B hlpA
71042106516016546123012prf: 2517386AThermus aquaticus dnaE30.257.51101DNA polymerase III
7114211654676655122447gp: SCE126_11Streptomyces coelicolor A3(2)41.562.3159hypothetical protein
SCE126.11
71242126551226565341413gp: SCE9_1Streptomyces coelicolor A3(2)26.156.0468hypothetical membrane protein
SCE9.01
7134213655834655097738
7144214656547657215669pir: C70884Mycobacterium tuberculosis50.376.4203transcriptional repressor
H37Rv Rv2788 sirR
7154215658002657205798gp: SCG8A_5Streptomyces coelicolor A3(2)34.961.7264hypothetical protein
SCG8A.05c
7164216658005658142138
7174217658155658928774pir: C69459Archaeoglobus fulgidus AF167642.571.8245transcriptional regulator (Sir2 family)
7184218658933659424492gp: SC5H1_34Streptomyces coelicolor A3(2)45.278.3157hypothetical protein
SC5H1.34
7194219659543660538996gp: CDU02617_1Corynebacterium diphtheriae31.162.2357iron-regulated lipoprotein precursor
irp1
7204220661120660650471pir: E70971Mycobacterium tuberculosis62.986.1151rRNA methylase
H37Rv Rv3366 spoU
7214221661166662017852pir: C70970Mycobacterium tuberculosis70.987.4278methylenetetrahydrofolate
H37Rv Rv3356c folDdehydrogenase
7224222662120662374255gp: MLCB1779_8Mycobacterium leprae31.376.380hypothetical membrane protein
MLCB1779.16c
72342236637616623821380gp: SC66T3_18Streptomyces coelicolor A3(2)34.063.2489hypothetical protein
SC66T3.18c
7244224665088664126963
72542256663136651831131gp: AF052652_1Corynebacterium glutamicum99.599.5379homoserine O-acetyltransferase
metA
72642266677706664601311prf: 2317335ALeptospira meyeri metY49.776.2429O-acetylhomoserine sulfhydrylase
72742276682646704652202sp: CSTA_ECOLIEscherichia coli K12 cstA53.978.4690carbon starvation protein
7284228670053669445609
7294229670472670672201sp: YJIX_ECOLIEscherichia coli K12 yjiX40.066.050hypothetical protein
7304230671653671045609
7314231671700672653954pir: C70539Mycobacterium tuberculosis71.086.4317hypothetical protein
H37Rv Rv1130
7324232672665673576912prf: 1902224AStreptomyces hygroscopicus41.676.2281carboxy phosphoenolpyruvate
mutase
73342336736086747561149sp: CISY_MYCSMMycobacterium smegmatis56.181.3380citrate synthase
ATCC 607 gltA
7344234673639672710930
7354235674990674799192sp: YNEC_ECOLIEscherichia coli K12 yneC34.062.353hypothetical protein
7364236675175675846672
73742376761226750821041sp: MDH_METFEMethanothermus fervidus V24S37.667.5338L-malate dehydrogenase
mdh
7384238676937676218720prf: 2514353LBacillus stearothermophilus T-626.162.8226regulatory protein
uxuR
7394239677748677047702
7404240681027680131897sp: VIUB_VIBCHVibrio cholerae OGAWA 39525.454.2284vibriobactin utilization protein
viuB
7414241681846681040807gp: AF176902_3Corynebacterium diphtheriae55.485.1269ABC transporter ATP-binding protein
irp1D
74242426829046818461059gp: AF176902_2Corynebacterium diphtheriae56.386.4339ABC transporter
irp1C
7434243683866682871996gp: AF176902_1Corynebacterium diphtheriae63.088.2330ABC transporter
irp1B
74442446849256838761050gp: CDU02617_1Corynebacterium diphtheriae53.182.3356iron-regulated lipoprotein precursor
irp1
74542456851096863801272prf: 2202262AStreptomyces venezuelae cmlv32.269.6395chloramphenicol resistance protein
7464246686435687346912prf: 2222220BPseudomonas aeruginosa crc30.458.1303catabolite repression control protein
7474247687351688007657sp: YICG_HAEINHaemophilus influenzae Rd56.285.8219hypothetical protein
HI1240
7484248688141688335195
7494249689890688916975
7504250690696689917780gp: AF109162_3Corynebacterium diphtheriae45.173.8244ferrichrome ABC transporter
hmuV
75142516917226907061017pir: S54438Yersinia enterocolitica hemU38.769.1346hemin permease
75242526918826929161035sp: SYW_ECOLIEscherichia coli K12 trpS54.479.8331tryptophanyl-tRNA synthetase
75342536930286941101083sp: YHJD_ECOLIEscherichia coli K12 yhjD37.172.3278hypothetical protein
7544254694172695074903
75542556962136950771137sp: DACD_SALTYSalmonella typhimurium LT230.957.5301penicillin-binding protein 6B
dacDprecursor
75642566979956967691227pir: F70842Mycobacterium tuberculosis34.170.7417hypothetical protein
H37Rv Rv3311
7574257698922698065858gp: SC6G10_8Streptomyces coelicolor A3(2)29.452.6323hypothetical protein
SC6G10.08c
7584258699072699266195
7594259699272698922351
7604260699281699913633sp: UPP_LACLALactococcus lactis upp46.472.3209uracil phosphoribosyltransferase
7614261699998700381384gp: SC1A2_11Streptomyces coelicolor A3(2)41.666.277bacterial regulatory protein, lacl
SC1A2.11family
76242627020817032621182pir: H70841Mycobacterium tuberculosis51.480.5385N-acyl-L-amino acid amidohydrolase
H37Rv Rv3305c amiAor peptidase
76342637021087003841725sp: MANB_MYCPIMycoplasma pirum BER manB22.153.8561phosphomannomutase
76442647034057048111407sp: DLDH_HALVOHalobacterium volcanii ATCC31.665.0468dihydrolipoamide dehydrogenase
29605 lpd
76542657052117086303420prf: 2415454ACorynebacterium glutamicum100.0100.01140pyruvate carboxylase
strain21253 pyc
7664266708839709708870sp: YD24_MYCTUMycobacterium tuberculosis26.260.1263hypothetical protein
H37Rv Rv1324
7674267709793710278486gp: SCF11_30Streptomyces coelicolor A3(2)30.766.9127hypothetical protein
SCF11.30
76842687116057105201086pir: B69760Bacillus subtilis 168 yciC44.669.0381hypothetical protein
7694269711724712647924sp: TRXB_BACSUBacillus subtilis IS58 trxB24.659.3305thioredoxin reductase
77042707127387142311494sp: PRPD_SALTYSalmonella typhimurium LT224.049.5521PrpD protein for propionate
prpDcatabolism
7714271714258715145888prf: 1902224AStreptomyces hygroscopicus42.574.5278carboxy phosphoenolpyruvate
mutase
7724272714757714380378PIR: E72779Aeropyrum pernix K1 APE022339.047.096hypothetical protein
77342737151027162831182sp: CISY_MYCSMMycobacterium smegmatis54.678.9383citrate synthase
ATCC 607 gltA
7744274716660716286375
77542757180097166871323pir: B70539Mycobacterium tuberculosis40.872.6456hypothetical protein
H37Rv Rv1129c
7764276718105718350246
77742777186587200161359
7784278721449720547903sp: THTR_CORGLCorynebacterium glutamicum100.0100.0225thiosulfate sulfurtransferase
ATCC 13032 thtR
77942797217777228411065gp: CJ11168X1_62Campylobacter jejuni Cj006961.179.8352hypothetical protein
7804280723338722925414gp: MLCB4_16Mycobacterium leprae51.176.7133hypothetical protein
MLCB4.27c
78142817234127255592148pir: G70539Mycobacterium tuberculosis35.163.4718hypothetical membrane protein
H37Rv Rv1565c
7824282726462725872591sp: YCEF_ECOLIEscherichia coli K12 yceF31.866.2192hypothetical protein
7834283726715726470246prf: 2323363CFMycobacterium leprae B1308-33.369.863hypothetical protein
C3-211
78442847283527267421611gp: AB018531_2Corynebacterium glutamicum99.8100.0537detergent sensitivity rescuer or
AJ11060 dtsR2carboxyl transferase
78542857303247286961629pir: JC4991Corynebacterium glutamicum99.6100.0543detergent sensitivity rescuer or
AJ11060 dtsR1carboxyl transferase
7864286730436731299864sp: BIRA_ECOLIEscherichia coli K12 birA28.761.8293bifunctional protein (biotin synthesis
repressor and biotin acetyl-CoA
carboxylase ligase)
7874287731312731797486pir: G70979Mycobacterium tuberculosis23.058.8165hypothetical membrane protein
H37Rv Rv3278c
78842887318577330171161sp: PURK_CORAMCorynebacterium69.083.83945′-phosphoribosyl-5-amino-4-
ammoniagenes ATCC 6872imidasol carboxylase
purK
78942897330727349431872sp: KUP_ECOLIEscherichia coli K12 kup41.173.6628K+-uptake protein
7904290733797733183615
7914291734984735340357
7924292735402735896495sp: PUR6_CORAMCorynebacterium85.793.21475′-phosphoribosyl-5-amino-4-
ammoniagenes ATCC 6872imidasol carboxylase
purE
7934293735899736351453gp: APU33059_5Actinosynnema pretiosum36.260.5152hypothetical protein
7944294736413737204792gp: SCF43A_36Streptomyces coelicolor A3(2)42.870.6255hypothetical protein
SCF43A.36
79542957385297372161314sp: NTAA_CHEHEChelatobacter heintzii ATCC43.273.0426nitrilotriacetate monooxygenase
29600 ntaA
79642967401727386731500pir: A69426Archaeoglobus fulgidus 23.452.5303transposase (ISA0963-5)
7974297741016740228789sp: DHG2_BACMEBacillus megaterium IAM 103031.364.8256glucose 1-dehydrogenase
gdhII
7984298741397741765369pir: A72258Thermotoga maritima MSB829.268.896hypothetical membrane protein
TM1408
7994299741854742195342
8004300742384741818567sp: YWJB_BACSUBacillus subtilis 168 ywjB28.666.3175hypothetical protein
8014301742409742828420gp: SCJ9A_21Streptomyces coelicolor A3(2)35.976.8142hypothetical protein
SCJ9A.21
8024302743052742831222
8034303743900743067834prf: 2406355CThermococcus litoralis malG42.475.3271trehalose/maltose-binding protein
80443047449317439001032prf: 2406355BThermococcus litoralis malF37.370.3306trehalose/maltose-binding protein
8054305745513745046468
80643067468937456221272prf: 2406355AThermococcus litoralis malE30.962.4417trehalose/maltose-binding protein
8074307748020748442423
8084308748026747031996prf: 2308356AStreptomyces reticuli msiK57.273.9332ABC transporter ATP-binding protein
(ABC-type sugar transport protein)
or celloblose/maltose transport
protein
8094309748446748814369
81043107536857488864800pir: B75633Deinococcus radiodurans R125.149.91783RNA helicase
DRB0135
8114311757063757434372
81243127573957536973699
8134313758262757630633pir: E70978Mycobacterium tuberculosis31.759.2240hypothetical protein
H37Rv Rv3268
81443147607967583642433pir: C71929Helicobacter pylori J99 jhp046230.062.5720hypothetical protein
81543157624687609061563sp: UVRD_ECOLIEscherichia coli K12 uvrD20.741.1701DNA helicase II
8164316762497762853357
8174317762730763122393
8184318762977762582396
8194319768191767367825
82043207694437632376207pir: T36671Streptomyces coelicolor22.445.82033RNA helicase
SCH5.13
82143217741427695474596pir: T08313Halobacterium sp. NRC-124.453.2698hypothetical protein
plasmid pNRC100 H1130
82243227770357741502886sp: HEPA_ECOLIEscherichia coli K12 hepA23.148.6873RNA polymerase associated protein
(ATP-dependent helicase)
82343237787117771581554pir: D70978Mycobacterium tuberculosis45.571.4527hypothetical protein
H37Rv Rv3267
8244324779014779910897gp: AF187550_1Mycobacterium smegmatis56.477.9289dTDP-Rha: a-D-GlcNAc-
mc2155 wbbLdiphosphoryl polyprenol, a-3-L-
rhamnosyl transferase
82543257801287811711044sp: MPG1_YEASTSaccharomyces cerevisiae29.866.9353mannose-1-phosphate
YDL055C MPG1guanylyltransferase
8264326781468781875408gp: AF164439_1Mycobacterium smegmatis73.481.994regulatory protein
whmD
8274327782617782162456pir: B70847Mycobacterium tuberculosis48.974.8139hypothetical protein
H37Rv Rv3259
8284328782712783101390gp: SCE34_11Streptomyces coelicolor A3(2)51.571.3136hypothetical protein
SCE34.11c
82943297831847845571374sp: MANB_SALMOSalmonella montevideo M4038.066.3460phosphomannomutase
manB
83043307846357856391005pir: B70594Mycobacterium tuberculosis31.256.3327hypothetical protein
H37Rv Rv3256c
83143317856437868241182sp: MANA_ECOLIEscherichia coli K12 manA36.966.2420mannose-6-phosphate isomerase
8324332786896787045150
8334333787624787983360
8344334787733787170564prf: 1804279KEnterococcus faecalis plasmid35.657.8180pheromone-responsive protein
pCF10 prgC
8354335788196788546351
83643367886727900931422sp: SAHH_TRIVATrichomonas vaginalis WAA3859.083.0476S-adenosyl-L-homocysteine
hydrolase
8374337789426788719708
8384338789721789002720
8394339790096790704609sp: KTHY_ARCFUArchaeoglobus fulgidus VC-1625.856.0209thymidylate kinase
AF0061
8404340790732791409678prf: 2214304AMycobacterium tuberculosis73.790.6224two-component system response
H37Rv Rv3246c mtrAregulator
8414341791421790738684
84243427915127930081497prf: 2214304BMycobacterium tuberculosis53.178.9484two-component system sensor
H37Rv Rv3245c mtrBhistidine kinase
84343437930087947111704pir: F70592Mycobacterium tuberculosis29.665.6595lipoprotein
H37Rv Rv3244c lpqB
8444344794714795301588pir: D70592Mycobacterium tuberculosis38.072.8213hypothetical protein
H37Rv Rv3242c
8454345795447795292156
8464346795448796110663sp: RR30_SPIOLSpinacia oleracea CV rps2234.561.620330S ribosomal protein or chloroplast
precursor
84743477962507987842535gsp: R74093Brevibacterium flavum99.199.6845preprotein translocase SecA subunit
(Corynebacterium glutamicum)
MJ-233 secA
8484348799020799691672
8494349799697800200504pir: A70591Mycobacterium tuberculosis47.178.8170hypothetical protein
H37Rv Rv3231c
8504350801194800208987pir: F70590Mycobacterium tuberculosis64.682.9322hypothetical protein
H37Rv Rv3228
85143518026028011901413gp: AF114233_1Corynebacterium glutamicum99.099.04615-enolpyruvylshikimate 3-phosphate
ASO19 aroAsynthase
8524352802649803128480pir: D70590Mycobacterium tuberculosis38.363.9180hypothetical protein
H37Rv Rv3226c
8534353802687802565123GP: AF114233_1Corynebacterium glutamicum100.0100.0235-enolpyruvylshikimate 3-phosphate
synthase
85443548042408031311110pir: G70506Mycobacterium tuberculosis21.642.4380hypothetical protein
H37Rv Rv0336
8554355804408805025618prf: 2515333DMycobacterium tuberculosis61.287.2188RNA polymerase sigma factor
sigH
8564356805792805535258pir: D70596Mycobacterium tuberculosis78.696.484regulatory protein
H37Rv Rv3219 whiB1
8574357806318806737420pir: B70596Mycobacterium tuberculosis33.365.1129hypothetical protein
H37Rv Rv3217c
85843588079398067401200pir: E70595Mycobacterium tuberculosis29.662.2415hypothetical protein
H37Rv Rv3212
85943598092178079461272sp: DEAD_KLEPNKlebsiella pneumoniae CG4337.364.0458DEAD box ATP-dependent RNA
deaDhelicase
8604360809286809510225
8614361809549810394846pir: H70594Mycobacterium tuberculosis46.469.8291hypothetical protein
H37Rv Rv3207c
8624362810405811163759pir: F70594Mycobacterium tuberculosis37.065.9249hypothetical protein
H37Rv Rv3205c
86343638111708142173048pir: G70951Mycobacterium tuberculosis23.948.91155ATP-dependent DNA helicase
H37Rv Rv3201c
8644364812165811386780
86543658142048174223219pir: G70951Mycobacterium tuberculosis41.465.71126ATP-dependent DNA helicase
H37Rv Rv3201c
86643668155418142101332
86743678175198185231005sp: Y13B_METJAMethanococcus jannaschii JAL-26.264.2302potassium channel
1 MJ0138.1.
8684368818523819236714pir: E70951Mycobacterium tuberculosis30.458.3230hypothetical protein
H37Rv Rv3199c
86943698192548212872034sp: UVRD_ECOLIEscherichia coli K12 uvrD32.658.8660DNA helicase II
8704370822079822669591
8714371822105821290816pir: B70951Mycobacterium tuberculosis26.849.3280hypothetical protein
H37Rv Rv3196
8724372822789823391603
87343738241258226801446pir: A70951Mycobacterium tuberculosis42.876.4474hypothetical protein
H37Rv Rv3195
87443748241908252391050pir: H70950Mycobacterium tuberculosis43.474.9350hypothetical protein
H37Rv Rv3194
8754375825916825242675
8764376826517825996522
87743778266168295702955pir: G70950Mycobacterium tuberculosis47.273.51023hypothetical protein
H37Rv Rv3193c
87843788309858296271359gp: AE001938_5Deinococcus radiodurans34.357.7463regulatory protein
DR0840
8794379831021831971951sp: ER1_HEVBRHevea brasiliensis laticifer er167.489.0301ethylene-inducible protein
8804380831922831578345PIR: F72782Aeropyrum pernix K1 APE024749.053.081hypothetical protein
8814381831971832570600sp: YAAE_BACSUBacillus subtilis 168 yaaE40.873.6201hypothetical protein
8824382833157832795363
88343838335728346331062pir: TRYXB4Lysobacter enzymogenes ATCC26.744.4408alpha-lytic proteinase precursor
29487
8844384834888835388501
8854385835253835837585pir: S03722Neurospora intermedia LaBelle-25.051.4208DNA-directed DNA polymerase
1b mitochondrion plasmid
88643868373128388921581sp: CSP1_CORGLCorynebacterium glutamicum27.051.5363major secreted protein PS1 protein
(Brevibacterium flavum) ATCCprecursor
17965 csp1
8874387838925839353429
8884388839630840139510
8894389840431840210222
8904390840745840437309
8914391842296841517780prf: 2207273HStreptomyces alboniger pur351.874.9255monophosphatase
8924392843124842306819gp: U70376_9Streptomyces flavopersicus33.759.3243myo-inositol monophosphatase
spcA
89343938432578443601104sp: RF2_STRCOStreptomyces coelicolor A3(2)68.088.6359peptide chain release factor 2
prfB
8944394844495845181687pir: E70919Mycobacterium tuberculosis70.491.2226cell division ATP-binding protein
H37Rv Rv3102c ftsE
8954395845105844842264PIR: G72510Aeropyrum pernix K1 APE206143.054.072hypothetical protein
8964396845198846097900pir: D70919Mycobacterium tuberculosis40.574.8301cell division protein
H37Rv Rv3101c ftsX
8974397846137846628492sp: SMPB_ECOLIEscherichia coli K12 smpB43.575.9145small protein B (SSRA-binding
protein)
8984398846632846982351sp: YEAO_ECOLIEscherichia coli K12 yeaO44.073.3116hypothetical protein
8994399846805846269537
9004400847727848026300
9014401848122847718405
9024402849323848499825sp: VIUB_VIBCHVibrio cholerae OGAWA 39526.852.9272vibriobactin utilization protein
viuB
9034403850243849326918prf: 2510361AStaphylococcus aureus sirA29.558.3319Fe-regulated protein
9044404850999850412588gp: MLCB1243_5Mycobacterium leprae36.171.2191hypothetical membrane protein
MLCB1243.07
90544058513518523641014sp: FATB_VIBANVibrio anguillarum 775 fatB27.761.5325ferric anguibactin-binding protein
precursor
9064406852618853616999pir: B69763Bacillus subtilis 168 yclN39.380.8313ferrichrome ABC transporter
(permease)
9074407853783854724942pir: C69763Bacillus subtilis 168 yclO35.676.0312ferrichrome ABC transporter
(permease)
9084408854724855476753pir: D69763Bacillus subtilis 168 yclP48.482.0250ferrichrome ABC transporter (ATP-
binding protein)
9094409860224860078147PIR: F81737Chlamydia muridarum Nigg66.072.048hypothetical protein
TC0129
9104410860745860473273GSP: Y35814Chlamydia pneumoniae61.066.084hypothetical protein
91144118615448627521209pir: S66270Rattus norvegicus (Rat)33.564.9442kynurenine
aminotransferase/glutamine
transaminase K
9124412863391862753639
91344138650668633961671sp: RA25_YEASTSaccharomyces cerevisiae30.762.3613DNA repair helicase
S288C YIL143C RAD25
91444148673178651192199pir: F70815Mycobacterium tuberculosis36.165.2764hypothetical protein
H37Rv Rv0862c
9154415867353867571219pir: G70815Mycobacterium tuberculosis44.062.057hypothetical protein
H37Rv Rv0863
9164416867788868630843
9174417868399867803597prf: 2420502AMicrococcus luteus rpf39.464.7198resuscitation-promoting factor
9184418868938869318381prf: 2320271ALactococcus lactis cspB42.675.461cold shock protein
9194419869903869379525gp: MLCB57_11Mycobacterium leprae28.358.5159hypothetical protein
MLCB57.27c
9204420870691869918774gp: AE001874_1Deinococcus radiodurans41.867.8273glutamine cyclotransferase
DR0112
9214421871419870721699
9224422871523871660138
92344238717388732101473gp: SC6C5_9Streptomyces coelicolor A3(2)43.679.3477permease
SC6C5.09
9244424872927872016912
9254425873213874040828sp: TSNR_STRAZStreptomyces azureus tsnR27.951.7319rRNA(adenosine-2′-O-)-
methyltransferase
9264426874944874069876
9274427875883874951933sp: YZ11_MYCTUMycobacterium tuberculosis32.655.1316hypothetical protein
H37Rv Rv0883c
92844288771128759851128pir: S71439Bacillus circulans ATCC 2178321.952.9374phosphoserine transaminase
92944298811148796421473sp: ACCD_ECOLIEscherichia coli K12 accD36.069.5236acetyl-coenzyme A carboxylase
carboxy transferase subunit beta
9304430881647881985339gp: SCI8_8Streptomyces coelicolor A3(2)51.580.6103hypothetical protein
SCI8.08c
93144318819958836471653pir: JC2382Pseudomonas fluorescens26.458.1549sodium/proline symporter
9324432883726884541816
9334433885388884549840pir: A70657Mycobacterium tuberculosis49.077.4243hypothetical protein
H37Rv Rv2525c
93444348856728945788907pir: S55505Corynebacterium63.183.43026fatty-acid synthase
ammoniagenes fas
9354435894703895191489
9364436895408895593186
93744378966428955961047prf: 2317335BLeptospira meyeri metX29.059.7335homoserine O-acetyltransferase
9384438897144896719426
9394439897423897689267
9404440897963897727237gp: AE002044_8Deinococcus radiodurans43.672.662glutaredoxin
DR2085
9414441898434897979456prf: 2408256AMycobacterium avium folA38.062.0171dihydrofolate reductase
9424442899231898434798sp: TYSY_ECOLIEscherichia coli K12 thyA64.888.9261thymidylate synthase
9434443900008899253756sp: CYSQ_ECOLIEscherichia coli K12 cysQ32.256.4202ammonium transporter
94444449000439046024560gp: SC7C7_16Streptomyces coelicolor A3(2)47.468.11715ATP dependent DNA helicase
SC7C7.16c
9454445904615905382768sp: FPG_SYNENSynechococcus elongatus29.251.0298formamidopyrimidine-DNA
naegeli mutMglycosidase
9464446905389905796408pir: F70816Mycobacterium tuberculosis55.586.7128hypothetical protein
H37Rv Rv0870c
9474447906391905792600sp: APL_LACLALactococcus lactis MG1363 apl38.871.9196alkaline phosphatase
94844489077319065591173pir: T36776Streptomyces coelicolor A3(2)33.867.0403integral membrane transporter
SCI28.06c
9494449908612909328717
95044509093789077591620pir: NUECEscherichia coli JM101 pgi52.477.0557glucose-6-phosphate isomease
95144519106969095211176pir: G70506Mycobacterium tuberculosis24.652.3195hypothetical protein
H37Rv Rv0336
9524452910843911223381
9534453911163910855309sp: YT26_MYCTUMycobacterium tuberculosis59.085.978hypothetical protein
H37Rv Rv0948c
95444549112269135142289sp: PCRA_BACSTBacillus stearothermophilus46.173.1763ATP-dependent helicase
NCA 1503 pcrA
95544559156999134772223gp: SCE25_30Streptomyces coelicolor A3(2)21.848.6885ABC transporter
SCE25.30
9564456916364915699666prf: 2420410PBacillus subtilis 168 yvrO43.871.4217ABC transporter
9574457916874916368507
9584458917680916970711pir: D70716Mycobacterium tuberculosis43.673.3236peptidase
H37Rv Rv0950c
95944599179289193521425sp: YT19_MYCTUMycobacterium tuberculosis31.160.8434hypothetical protein
H37Rv Rv0955
9604460918054917827228
9614461919330919956627gp: AB003159_2Corynebacterium64.686.21895′-phosphoribosylglycinamide
ammoniagenes purNformyltransferase
96244629199679215261560gp: AB003159_3Corynebacterium74.587.85255′-phosphoribosyl-5-aminoimidazole-
ammoniagenes purH4-carboxamide formyltransferase
9634463921594922412819gp: CGL133719_3Corynebacterium glutamicum100.0100.0217citrate lyase (subunit)
ATCC 13032 citE
9644464923061922396666gp: CGL133719_2Corynebacterium glutamicum100.0100.0222repressor of the high-affinity (methyl)
ATCC 13032 amtRammonium uptake system
9654465923464923138327gp: CGL133719_1Corynebacterium glutamicum100.0100.0109hypothetical protein
ATCC 13032 yjcC
9664466923661923981321
9674467924407924159249sp: RR18_CYAPACyanophora paradoxa rps1852.276.16730S ribosomal protein S18
9684468924727924425303sp: RS14_ECOLIEscherichia coli K12 rpsN54.080.010030S ribosomal protein S14
9694469924895924734162sp: RL33_ECOLIEscherichia coli K12 rpmG55.183.74950S ribosomal protein L33
9704470925134924901234pir: R5EC28Escherichia coli K12 rpmB52.081.87750S ribosomal protein L28
97144719269359253251611pir: B70033Bacillus subtilis 168 yvdB34.471.1529transporter (sulfate transporter)
9724472927242926931312prf: 2420312AStaphylococcus aureus zntR37.577.580Zn/Co transport repressor
9734473927474927737264sp: RL31_HAEDUHaemophilus ducreyi rpmE37.265.47850S ribosomal protein L31
9744474927752927922171gp: SC51A_14Streptomyces coelicolor A3(2)60.078.25550S ribosomal protein L32
SCF51A.14
9754475927785927339447
9764476928117928812696sp: COPR_PSESMPseudomonas syringae copR48.073.6227copper-inducible two-component
regulator
97744779288849302481365sp: BAES_ECOLIEscherichia coli K12 baeS24.460.1484two-component system sensor
97844789304109316481239pir: S45229Escherichia coli K12 htrA33.359.9406proteinase DO precursor
9794479931706932290585sp: CNX1_ARATHArabidopsis thaliana CV cnx127.754.3188molybdopterin biosynthesis cnx1
protein (molybdenum cofactor
biosynthesis enzyme cnx1)
9804480932290932487198
9814481932974932570405sp: MSCL_MYCTUMycobacterium tuberculosis50.477.1131large-conductance
H37Rv Rv0985c mscLmechanosensitive channel
9824482933710933060651pir: A70601Mycobacterium tuberculosis28.660.0210hypothetical protein
H37Rv Rv0990
9834483934302933733570pir: JC4389Homo sapiens MTHFS25.159.71915-formyltetrahydrofolate cyclo-ligase
9844484934423935319897pir: JC4985Xanthomonas campestris42.268.9296UTP—glucose-1-phosphate
uridylyltransferase
98544859353519366071257prf: 2403296BArthrobacter nicotinovorans31.862.6390molybdopterin biosynthesis protein
moeA
9864486936615937274660sp: RIMJ_ECOLIEscherichia coli K12 rimJ29.054.9193ribosomal-protein-alanine N-
acetyltransferase
98744879373829384011020pir: G70601Mycobacterium tuberculosis30.354.8367hypothetical membrane protein
H37Rv Rv0996
98844889384279396261200sp: CYNX_ECOLIEscherichia coli K12 cynX26.662.4380cyanate transport protein
98944899392179377991419
9904490939686940090405sp: YG02_HAEINHaemophilus influenzae Rd32.160.6137hypothetical membrane protein
HI1602
9914491940041940754714sp: Y05C_MYCTUMycobacterium tuberculosis25.359.6225hypothetical membrane protein
H37Rv Rv0093c
99244929407599419251167sp: CDAS_BACSHBacillus sphaericus E-24426.853.6444cyclomaltodextrinase
CDase
99344939439409423811560pir: E70602Mycobacterium tuberculosis43.075.2488hypothetical membrane protein
H37Rv
9944494944009944833825sp: Y19J_MYCTUMycobacterium tuberculosis54.078.3272hypothetical protein
H37Rv Rv1003
99544959468409486691830sp: SYM_METTHMethanobacterium33.866.7615methionyl-tRNA synthetase
thermoautotrophicum Delta H
MTH587 metG
99644969487919508392049prf: 1306383AEscherichia coli recQ26.249.0741ATP-dependent DNA helicase
9974497951460950828633pir: B69206Methanobacterium27.653.3210hypothetical protein
thermoautotrophicum Delta H
MTH796
99844989529919518341158sp: YXAG_BACSUBacillus subtilis 168 yxaG30.059.0363hypothetical protein
9994499953573953043531
10004500953973954266294gp: AF029727_1Enterococcus faecium33.059.694transposase
10014501954277954753477pir: TQECI3Escherichia coli K1241.767.6139transposase
10024502954941955354414gp: AF052055_1Brevibacterium linens tnpA73.288.4112transposase subunit
10034503955911956774864
100445049573989556861713prf: 2014253AEEscherichia coli dld46.475.6565D-lactate dehydrogenase
10054505958683957844840sp: MTK1_KLEPNKlebsiella pneumoniae OK830.862.8231site-specific DNA-methyltransferase
kpnIM
10064506959403959185219
10074507960081960374294gp: AF029727_1Enterococcus faecium33.059.694transposase
10084508960385960861477pir: TQECI3Escherichia coli K1241.767.6139transposase
10094509961297961653357sp: YJ94_MYCTUMycobacterium tuberculosis62.684.691transcriptional regulator
H37Rv Rv1994c
10104510961629962249621prf: 2514367AStaphylococcus aureus cadD31.766.8205cadmium resistance protein
10114511961662961321342
10124512962809963639831pir: C70603Mycobacterium tuberculosis46.470.7263hypothetical protein
H37Rv Rv1008
101345139638649649341071pir: D70603Mycobacterium tuberculosis34.863.5362hypothetical protein
H37Rv Rv1009 rpf
10144514964974965852879sp: KSGA_ECOLIEscherichia coli K12 ksgA34.365.3265dimethyladenosine transferase
10154515965852966784933pir: F70603Mycobacterium tuberculosis42.567.0315isopentenyl monophosphate kinase
H37Rv Rv1011
10164516966591965950642
101745179668289686601833pir: S47441Saccharopolyspora erythraea65.585.8478ABC transporter
ertX
10184518968667969458792sp: PDXK_ECOLIEscherichia coli K12 pdxK40.167.4242pyridoxine kinase
10194519969940969461480sp: YX05_MYCTUMycobacterium tuberculosis27.058.5159hypothetical protein
H37Rv Rv2874
10204520970029970349321gp: SCF1_2Streptomyces coelicolor A3(2)45.478.7108hypothetical protein
SCF1.02
10214521970418970738321gp: SCF1_2Streptomyces coelicolor A3(2)35.569.2107hypothetical protein
SCF1.02
10224522970864971823960gp: SCJ1_15Streptomyces coelicolor A3(2)64.888.1261regulator
SCJ1.15
10234523973035972244792sp: YXEH_BACSUBacillus subtilis 168 yxeH27.259.1276hypothetical protein
102445249731399741551017pir: E70893Mycobacterium tuberculosis35.670.9337enoyl-CoA hydratase
H37Rv echA9
10254525973957973304654
10264526974186974962777
102745279761769749651212
102845289763499777341386sp: CSP1_CORGLCorynebacterium glutamicum27.756.8440major secreted protein PS1 protein
(Brevibacterium flavum) ATCCprecursor
17965 csp1
10294529978378977800579gp: SCF56_6Streptomyces coelicolor A3(2)44.070.0100transcriptional regulator (tetR
SCF56.06family)
103045309807409783682373gp: SCE87_17Streptomyces coelicolor A3(2)42.670.0802membrane transport protein
SCE87.17c
10314531980993981490498sp: MENG_HAEINHaemophilus influenzae Rd38.275.8157S-adenosylmethionine: 2-
HI0508 menGdemethylmenaquinone
methyltransferase
10324532981622982287666
10334533982674982294381gp: NMA6Z2491_214Neisseria meningitidis NMA195329.863.6121hypothetical protein
103445349831009846501551pir: A70539Mycobacterium tuberculosis24.948.3482hypothetical protein
H37Rv Rv1128c
10354535984910985845936
103645369865109848641647pir: I59305Escherichia coli K12 prfC39.268.0546peptide-chain-release factor 3
103745379867399880071269prf: 2406311AMethylophilus methylotrophus42.872.8404amide-urea transport protein
fmdD
10384538988023988904882prf: 2406311BMethylophilus methylotrophus40.861.077amide-urea transport protein
fmdE
103945399889049899801077prf: 2406311CMethylophilus methylotrophus34.668.0234amide-urea transport protein
fmdF
10404540989980990705726sp: BRAF_PSEAEPseudomonas aeruginosa PAO37.970.0253high-affinity branched-chain amino
braFacid transport ATP-binding protein
10414541990716991414699sp: BRAG_PSEAEPseudomonas aeruginosa PAO35.269.1236high-affinity branched-chain amino
braGacid transport ATP-binding protein
10424542992028991417612sp: PTH_ECOLIEscherichia coli K12 pth39.070.6187peptidyl-tRNA hydrolase
104345439920589930801023sp: 2NPD_WILMRWilliopsis mrakii IFO 089525.254.03612-nitropropane dioxygenase
104445449935499946131065sp: G3P_ZYMMOStreptomyces roseofulvus gap39.572.8342glyceraldehyde-3-phosphate
dehydrogenase
10454545994474994106369GSP: Y75094Neisseria meningitidis54.061.051polypeptides predicted to be useful
antigens for vaccines and
diagnostics
10464546995375994845531sp: PTH_ECOLIEscherichia coli K12 pth38.563.2174peptidyl-tRNA hydrolase
10474547996126995527600pir: B70622Mycobacterium tuberculosis47.065.019450S ribosomal protein L25
H37Rv rplY
10484548996402996830429sp: LGUL_SALTYSalmonella typhimurium D2128.754.6143lactoylglutathione lyase
gloA
10494549997456996833624prf: 2516401BWBacillus cereus ATCC 1098738.962.5208DNA alkylation repair enzyme
alkD
10504550998440997466975sp: KPRS_BACCLBacillus subtilis prs44.079.1316ribose-phosphate
pyrophosphokinase
105145519999099984551455pir: S66080Bacillus subtilis gcaD42.071.9452UDP-N-acetylglucosamine
pyrophosphorylase
10524552100124210000161227
10534553100133210028641533sp: SUFI_ECOLIEscherichia coli K12 sufI30.861.7506sufI protein precursor
1054455410030131003930918sp: NODI_RHIS3Rhizobium sp. N33 nodI35.864.8310nodulation ATP-binding protein I
1055455510039531004783831pir: JN0850Streptomyces lividans ORF230.263.2272hypothetical membrane protein
10564556100482910060851257sp: UHPB_ECOLIEscherichia coli K12 uhpB24.648.4459two-component system sensor
histidine kinase
1057455710060891006697609prf: 2107255AStreptomyces peucetius dnrN36.667.3202two component transcriptional
regulator (luxR family)
1058455810069371006734204
10594559100699810081521155gp: SCF15_7Streptomyces coelicolor A3(2)31.564.5349hypothetical membrane protein
SCF15.07
10604560100862210100611440pir: S65587Streptomyces glaucescens strV28.657.0535ABC transporter
1061456110086861008534153
10624562101005710117901734pir: T14180Mycobacterium smegmatis exiT44.074.0573ABC transporter
10634563101376110117971965sp: GGT_ECOLIEscherichia coli K12 ggt32.458.6666gamma-glutamyltranspeptidase
precursor
1064456410140161014264249
1065456510148611014343519
1066456610149251015116192
1067456710156521016560909
1068456810156921015450243GPU: AF164956_23Corynebacterium glutamicum64.072.037transposase protein fragment
TnpNC
1069456910158521015145708gp: AF121000_8Corynebacterium glutamicum99.6100.0236transposase (IS1628 TnpB)
22243 R-plasmid pAG1 tnpB
1070457010165571017018462
1071457110178701017274597
1072457210180821018393312
1073457310184161019066651sp: TETC_ECOLIEscherichia coli tetR23.059.6183transcriptional regulator (TetR-
family)
10744574101909010227163627sp: MFD_ECOLIEscherichia coli mfd36.265.11217transcription/repair-coupling protein
10754575102061310193901224
1076457610213051021078228GSP: Y75301Neisseria gonorrhoeae48.069.076Neisserial polypeptides predicted to
be useful antigens for vaccines and
diagnostics
10774577102466610226991968sp: MDLB_ECOLIEscherichia coli mdlB31.362.7632multidrug resistance-like ATP-
binding protein, ABC-type transport
protein
10784578102639610246661731sp: YC73_MYCTUMycobacterium tuberculosis50.281.9574ABC transporter
H37Rv Rv1273c
10794579102888610265052382sp: YLI3_CORGLCorynebacterium glutamicum100.0100.0368hypothetical membrane protein
ATCC 13032 orf3
1080458010318851032181297
1081458110321961032780585sp: YABN_BACSUBacillus subtilis yabN33.457.4183hypothetical protein
1082458210331851032760426
1083458310336461033269378
1084458410339541034739786pir: A70623Mycobacterium tuberculosis46.568.9241lpqU protein
H37Rv Rv1022 lpqU
10854585103494910362231275sp: ENO_BACSUBacillus subtilis eno64.586.0422enolase (2-phosphoglycerate
dehydratase)(2-phospho-D-
glycerate hydro-lyase)
1086458610361591036016144PIR: B72477Aeropyrum pernix K1 APE245968.058.041hypothetical protein
1087458710363161036855540pir: C70623Mycobacterium tuberculosis31.955.0191hypothetical protein
H37Rv Rv1024
1088458810369001037445546pir: D70623Mycobacterium tuberculosis59.577.8153hypothetical protein
H37Rv Rv1025
1089458910374481038410963sp: GPPA_ECOLIEscherichia coli gppA25.255.0329guanosine pentaphosphatase or
exopolyphosphatase
1090459010374811036498984
1091459110396501038721930sp: THD2_ECOLIEscherichia coli tdcB30.364.7314threonine dehydratase
1092459210397831039977195
1093459310399961040325330
1094459410404941040682189pir: B72287Thermotoga maritima MSB846.374.156hypothetical protein
1095459510409251041917993sp: RHAR_ECOLIEscherichia coli rhaR24.855.8242transcription activator of L-rhamnose
operon
1096459610420271042842816pir: F70893Mycobacterium tuberculosis57.880.1282hypothetical protein
H37Rv Rv1072
1097459710432361042850387
1098459810437471043298450gp: SCF55_39Streptomyces coelicolor A3(2)30.057.1140hypothetical protein
SCF55.39
1099459910442951043774522sp: GREA_ECOLIEscherichia coli greA35.060.1143transcription elongation factor
1100460010449591044477483pir: G70894Mycobacterium tuberculosis34.372.1140hypothetical protein
H37Rv Rv1081c
1101460110451581046030873pir: S44952Streptomyces lincolnensis lmbE31.756.3300lincomycin-production
1102460210460731046390318
11034603104661010477071098sp: AROG_CORGLCorynebacterium glutamicum99.299.53673-deoxy-D-arabino-heptulosonate-7-
aroGphosphate synthase
1104460410474521046820633
1105460510478271048501675sp: YARF_CORGLCorynebacterium glutamicum96.097.397hypothetical protein or undecaprenyl
CCRC18310pyrophosphate synthetase
1106460610483561048529174SP: YARF_CORGLCorynebacterium glutamicum100.0100.028hypothetical protein
(Brevibacterium flavum)
1107460710485251049043519
1108460810493851049068318
1109460910503621049427936sp: COAA_ECOLIEscherichia coli coaA53.979.9308pantothenate kinase
11104610105062410519251302gsp: R97745Brevibacterium flavum MJ-23399.5100.0434serine hydroxymethyl transferase
glyA
11114611105202110538801860sp: PABS_STRGRStreptomyces griseus pabS47.670.1898p-aminobenzoic acid synthase
1112461210538801054602723
1113461310548591055722864
1114461410550321054640393
1115461510557831056319537gp: A01504_1Alcaligenes faecalis ptcR30.358.8165phosphinothricin resistance protin
1116461610572001056322879sp: YBGK_ECOLIEscherichia coli ybgK30.359.0300hypothetical protein
11174617105757310586281056
1118461810578681057200669sp: YBGJ_ECOLIEscherichia coli ybgJ37.857.8225hypothetical protein
1119461910585981057843756sp: LAMB_EMENIEmericella nidulans lamB30.852.2276lactam utilization protein
1120462010592141058624591sp: YCSH_BACSUBacillus subtilis ycsH40.681.2165hypothetical membrane protein
1121462110592181059889672
1122462210593601059962603
1123462310601121060792681sp: YDHC_BACSUBacillus subtilis ydhC26.063.2204transcriptional regulator
11244624106086910621461278
11254625106362910622111419sp: FUMH_RATRattus norvegicus (Rat) fumH52.079.4456fumarate hydratase precursor
1126462610639361064424489gp: AF048979_1Rhodococcus erythropolis 32.765.4159NADH-dependent FMN
IGTS8 dszDoxydoreductase
1127462710647381064478261
1128462810652001064754447
1129462910658671065304564gp: SCAH10_16Streptomyces coelicolor A3(2)55.481.0184reductase
StAH10.16
11304630106608310675701488sp: SOXA_RHOSORhodococcus sp. IGTS8 soxA39.167.7443dibenzothiophene desulfurization
enzyme A
11314631106757010686491080sp: SOXC_RHOSORhodococcus sp. IGTS8 soxC25.851.3372dibenzothiophene desulfurization
enzyme C (DBT sulfur dioxygenase)
11324632106864910698451197sp: SOXC_RHOSORhodococcus sp. IGTS8 soxC28.961.6391dibenzothiophene desulfurization
enzyme C (DBT sulfur dioxygenase)
1133463310696921068913780
1134463410698081069119690
11354635106995910711341176gp: ECO237695_3Escherichia coli K12 ssuD45.373.1397FMNH2-dependent aliphatic
sulfonate monooxygenase
1136463610724411071479963sp: GLPX_ECOLIEscherichia coli K12 glpX44.375.7325glycerol metabolism
1137463710726761073245570pir: B70897Mycobacterium tuberculosis27.556.4211hypothetical protein
H37Rv Rv1100
11384638107524110733401902pir: H70062Bacillus subtilis ywmD31.366.1227hypothetical protein
1139463910753571075641285
1140464010755531075329225gp: SCH24_37Streptomyces coelicolor A3(2)36.678.182transmembrane efflux protein
SCH24.37
1141464110759091075667243sp: EX7S_ECOLIEscherichia coli K12 MG165540.367.762exodeoxyribonuclease small subunit
xseB
11424642107718310759331251sp: EX7L_ECOLIEscherichia coli K12 MG165530.055.6466exodeoxyribonuclease large subunit
xseA
1143464310772971078271975sp: LYTB_ECOLIEscherichia coli K12 lytB50.278.8311penicillin tolerance
1144464410777341077306429GSP: Y75421Neisseria gonorrhoeae33.047.0131polypeptides predicted to be useful
antigens for vaccines and
diagnostics
1145464510791461078319828
11464646108054010792211320sp: PERM_ECOLIEscherichia coli K12 perM26.363.9338permease
1147464710809651080786180
11484648108270810809721737sp: NTPR_RATRattus norvegicus (Rat) SLC6A730.361.4552sodium-dependent proline
ntpRtransporter
11494649108418310829511233sp: CSP1_CORGLCorynebacterium glutamicum29.960.0412major secreted protein PS1 protein
(Brevibacterium flavum) ATCCprecursor
17965 csp1
11504650108438010854621083sp: YYAF_BACSUBacillus subtilis yyaF70.188.6361GTP-binding protein
1151465110857911086087297sp: VAPI_BACNODichelobacter nodosus intA57.380.075virulence-associated protein
1152465210860961086917822sp: OTCA_PSEAEPseudomonas aeruginosa argF29.658.8301ornithine carbamoyltransferase
1153465310875441087044501sp: YKKB_BACSUBacillus subtilis 168 ykkB39.269.9143hypothetical protein
1154465410882931087664630gp: AF013288_1Mus musculus RDH433.860.61989-cis retinol dehydrogenase or
oxidoreductase
11554655108974010885351206sp: YIS1_STRCOStreptomyces coelicolor42.273.0396transposase/integrase (IS110)
SC3C8.10
11564656109017510932163042sp: YEGE_ECOLIEscherichia coli K12 yegE23.052.21153hypothetical membrane protein
1157465710939291094693765sp: NODC_RHIMERhizobium meliloti nodC22.847.1259N-acetylglucosaminyltransferase
1158465810946931094911219
1159465910950521095384333
1160466010956771095387291pir: S43613Corynebacterium glutamicum82.593.897transposase (insertion sequence
ATCC 31831IS31831)
1161466110960931095719375pir: JC4742Corynebacterium glutamicum79.294.4125transposase
(Brevibacterium lactofermentum)
ATCC 13869
1162466210963311096188144pir: JC4742Corynebacterium glutamicum87.595.848transposase
(Brevibacterium lactofermentum)
ATCC 13869
1163466310964711096331141
1164466410971111096746366
1165466510972291097726498
1166466610977501098592843sp: MORA_PSEPUPseudomonas putida M10 norA37.566.3264oxidoreductase or morpyine-6-
dehydrogenase (naloxone
reductase)
1167466710986091098929321sp: DC4C_ACICAAcinetobacter calcoaceticus33.363.91084-carboxymuconolactone
dc4cdecarboxlyase
1168466810990881099750663
1169466910992091099015195
1170467010997681099115654gp: AF058302_19Streptomyces roseofulvus frnS34.966.4146frenolicin gene cluster protein
involved in frenolicin biosynthetic
11714671109991711016531737gp: SPU59234_3Synechococcus sp. PCC 794248.178.5563biotin carboxylase
accC
1172467211020431102639597
1173467311026951103192498
1174467411031801103524345
1175467511039511104103153
1176467611049231105561639
11774677110605811041031956sp: YT15_MYCTUMycobacterium tuberculosis57.980.3655hypothetical protein
H37Rv Rv0959
11784678110738111060861296sp: BCHI_RHOSHRhodobacter sphaeroides ATCC27.752.6329magnesium chelatase subunit
17023 bchl
1179467911075601108201642gp: AMU73808_1Amycolatopsis methanolica pgm33.862.51602,3-PDG dependent
phosphoglycerate mutase
1180468011082011108905705pir: A70577Mycobacterium tuberculosis38.260.7262hypothetical protein
H37Rv Rv2133c
1181468111089931109754762gp: STMBCPA_1Streptomyces hygroscopicus29.459.3248carboxyphosphonoenolpyruvate
SF1293 BcpAphosphonomutase
11824682110979211114321641sp: TLRC_STRFRStreptomyces fradiae tlrC31.754.1593tyrosin resistance ATP-binding
protein
1183468311118201111425396sp: Y06C_MYCTUMycobacterium tuberculosis29.466.9136hypothetical protein
H37Rv Rv2923c
1184468411118891112230342sp: PHNA_ECOLIEscherichia coli K12 MG165555.082.0111alkylphosphonate uptake protein
phnA
1185468511129571112484474sp: YXAD_BACSUBacillus subtilis 168 yxaD32.162.7134transcriptional regulator
11864686111310211143191218gp: SPN7367_1Streptococcus pneumoniae22.659.4367multi-drug resistance efflux pump
pmrA
11874687111448611157931308pir: S43613Corynebacterium glutamicum99.599.8436transposase (insertion sequence
(Brevibacterium lactofermentum)IS31831)
ATCC 31831
11884688111690511158321074gp: RFAJ3152_2Ruminococcus flavefaciens43.973.4376cysteine desulphurase
cysteine desulphurase gene
1189468911177441116908837sp: NADC_MYCTUMycobacterium tuberculosis42.168.9283nicotinate-nucleotide
pyrophosphorylase
11904690111893211177511182pir: E69663Bacillus subtilis nadA49.377.6361quinolinate synthetase A
1191469111197271119086642gp: SC5B8_7Streptomyces coelicolor37.060.9235DNA hydrolase
SC5B8.07
1192469211202051120804600gp: AE001961_5Deinococcus radiodurans R123.454.7192hypothetical membrane protein
DR1112
1193469311214321120833600gp: SC3A7_8Streptomyces coelicolor36.066.4214hypothetical protein
SC3A7.08
1194469411218091121468342sp: YBDF_ECOLIEscherichia coli K12 MG165541.774.1108hypothetical protein
ybdF
1195469511226061121818789gp: AAA21740_1Escherichia coli K12 lplA30.160.7216lipoate-protein ligase A
1196469611230511123461411sp: PHNB_ECOLIEscherichia coli K12 phnB29.760.8148alkylphosphonate uptake protein
and C-P lyase activity
11974697112482611235341293sp: PCAK_PSEPUPseudomonas putida pcaK28.864.3420transmembrane transport protein or
4-hydroxybenzoate transporter
11984698112602011248361185sp: PHHY_PSEAEPseudomonas aeruginosa phhy40.868.6395p-hydroxybenzoate hydroxylase (4-
hydroxybenzoate 3-
monooxygenase)
1199469911264221127009588pir: A69859Bacillus subtilis 168 ykoE36.769.6191hypothetical membrane protein
12004700112701311283501338sp: YJJK_ECOLIEscherichia coli yjjK24.847.6532ABC transporter ATP-binding protein
1201470111283501129102753pir: G69858Bacillus subtilis 168 ykoC25.661.6250hypothetical membrane protein
1202470211291021129632531
12034703112965511307041050sp: CHAA_ECOLIEscherichia coli chaA33.369.0339Ca2+/H+ antiporter ChaA
1204470411307211131428708pir: C75001Pyrococcus abyssi Orsay28.457.6236hypothetical protein
PAB1341
1205470511321231131401723sp: YWAF_BACSUBacillus subtilis ywaF27.661.1221hypothetical membrane protein
12064706113447211321332340sp: UVRA_THETHThermus thermophilus unrA35.558.7946excinuclease ABC subunit A
1207470711345611135055495sp: TPX_MYCTUMycobacterium tuberculosis57.381.7164thioredoxin peroxidase
H37Rv tpx
1208470811354761135691216
12094709113683311350581776
1210471011378911136938954sp: YEDI_ECOLIEscherichia coli yedL39.972.0318hypothetical membrane protein
1211471111379601138859900gp: SCF76_2Streptomyces coelicolor A3(2)34.049.0282oxidoreductase or thiamin
biosynthesis protein
1212471211388801139245366
1213471311391961139492297
1214471411393571139617261
1215471511400211139635387
1216471611408611140028834sp: CTR2_PENVAPenaeus vannamei28.851.3271chymotrypsin BII
1217471711412451140901345sp: ARC2_ECOLIEscherichia coli43.272.1111arsenate reductase (arsenical pump
modifier)
12184718114127311424721200sp: YYAD_BACSUBacillus subtilis yyaD23.562.4340hypothetical membrane protein
1219471911430151142479537pir: F70559Mycobacterium tuberculosis43.571.4147hypothetical protein
H37Rv Rv1632c
1220472011437391143026714pir: F70555Mycobacterium tuberculosis35.862.9221hypothetical protein
H37Rv Rv1157c
12214721114411811460281911sp: TYPA_ECOLIEscherichia coli K12 typA46.376.7614GTP-binding protein (tyrosine
phsphorylated protein A)
12224722114609711476021506pir: F70874Mycobacterium tuberculosis27.954.9506hypothetical protein
H37Rv Rv1166
1223472311475921148461870pir: B70875Mycobacterium tuberculosis38.761.9315hypothetical protein
H37Rv Rv1170
1224472411484451148882438
1225472511489531149267315sp: FER_STRGRStreptomyces griseus fer78.691.3103ferredoxin [4Fe-4S]
12264726114927911503791101sp: AAT_BACSPBacillus sp: strain YM-2 aat25.952.9397aspartate aminotransferase
1227472711504081151028621
12284728115118611523701185
1229472911532631152373891gp: CGAJ4934_1Corynebacterium glutamicum100.0100.0229tetrahydrodipicolinate succinylase or
ATCC 13032 dapDsuccinylation of piperidine-2,6-
dicarboxylate
1230473011565371155875663
1231473111569021157669768pir: S60064Corynebacterium glutamicum100.0100.0211hypothetical protein
ATCC 13032 orf2
1232473211576941158524831gp: SCP8_4Streptomyces coelicolor A3(2)59.069.0273dihydropteroate synthase
dhpS
1233473311585241159252729gp: MLU15180_14Mycobacterium leprae u1756I45.773.1245hypothetical protein
1234473411592671159572306pir: G70609Mycobacterium tuberculosis31.367.799hypothetical protein
H37Rv Rv1209
1235473511596351159799165gsp: W32443Mycobacterium tuberculosis72.391.547antigen TbAAMK, useful in vaccines
for prevention or treatment of
tuberculosis
1236473611598651160728864sp: MYRA_MICGRMicromonospora griseorubida39.267.8286mycinamicin-resistance gene
myrA
12374737116223111607381494sp: SCRB_PEDPEPediococcus pentosaceus scrB23.551.0524sucrose-6-phosphate hydrolase
12384738116360511623791227sp: GLGA_ECOLIEscherichia coli K12 MG165524.751.3433ADPglucose—starch(bacterial
glgAglycogen) glucosyltransferase
12394739116370211649161215sp: GLGC_STRCOStreptomyces coelicolor A3(2)61.081.8400glucose-1-phosphate
glgCadenylyltransferase
1240474011656121164974639sp: MDMC_STRMYStreptomyces mycarofaciens25.862.493methyltransferase
MdmC
1241474111657461166384639sp: RPOE_ECOLIEscherichia coli rpoE27.357.2194RNA polymerase sigma factor
(sigma-24); heat shock and
oxidative stress
1242474211665761167067492
1243474311671101167577468pir: C70508Mycobacterium tuberculosis45.573.2112hypothetical protein
H37Rv Rv1224
12444744116871111675871125sp: MRP_ECOLIEscherichia coli mrp43.672.0257ATPase
1245474511693251168747579pir: B70509Mycobacterium tuberculosis60.483.8154hypothetical protein
H37Rv Rv1231c
12464746117061011693211290pir: C70509Mycobacterium tuberculosis49.877.0434hypothetical protein
H37Rv Rv1232c
1247474711706721171187516pir: A70952Mycobacterium tuberculosis57.987.1140hypothetical protein
H37Rv Rv1234
1248474811712061171871666
1249474911724621171869594
12504750117627111725013771prf: 2306367ACorynebacterium glutamicum99.499.812572-oxoglutarate dehydrogenase
AJ12036 odhA
12514751118004811763083741sp: MDR2_CRIGRCricetulus griseus (Chinese28.860.41288ABC transporter or multidrug
hamster) MDR2resistance protein 2 (P-glycoprotein
2)
1252475211808371180121717pir: H70953Mycobacterium tuberculosis31.772.1240hypothetical protein
H37Rv Rv1249c
1253475311816751180872804sp: AROE_ECOLIEscherichia coli aroE25.561.2255shikimate dehydrogenase
12544754118199311836031611sp: PNBA_BACSUBacillus subtilis pnbA35.764.7501para-nitrobenzyl esterase
1255475511836071184257651
1256475611842801185155876
1257475711857421185218525
12584758118582511870391215sp: TCR1_ECOLIEscherichia coli transposon27.161.4409tetracycline resistance protein
Tn1721 tetA
12594759118704311883891347sp: TCMA_STRGAStreptomyces glaucescens tcmA32.464.2444metabolite export pump of
tetracenomycin C resistance
1260476011898221190526705
12614761119062211883882235pir: S57636Catharanthus roseus metE45.272.27745-
methyltetrahydropteroyltriglutamate—
homocysteine S-methyltransferase
1262476211910871191542456
12634763119241011938071398gsp: Y29930Nocardia asteroides strain KGB155.279.5444thiophene biotransformation protein
1264476411938671194190324
1265476511941651195109945
1266476611959161195125792
12674767119597411976201647
1268476811976241197815192
12694769119954311979901554sp: CYDC_ECOLIEscherichia coli K12 MG165528.763.5526ABC transporter
cydC
12704770120107511995431533sp: CYDD_ECOLIEscherichia coli K12 MG165529.458.4551ABC transporter
cydD
1271477112020881201090999gp: AB035086_2Corynebacterium glutamicum92.093.0333cytochrome bd-type menaquinol
(Brevibacterium lactofermentum)oxidase subunit II
cydB
12724772120363212020941539gp: AB035086_1Corynebacterium glutamicum99.699.0512cytochrome bd-type menaquinol
(Brevibacterium lactofermentum)oxidase subunit I
cydA
12734773120618012039162265sp: YEJH_ECOLIEscherichia coli K12 MG165526.455.0402helicase
yejH
1274477412063161206657342
1275477512072231206831393sp: MUTT_PROVUProteus vulgaris mutT36.965.698mutator mutT protein ((7,8-dihydro-
8-oxoguanine-triphosphatase)(8-
oxo-dGTPase)(dGTP
pyrophosphohydrolase)
1276477612073741208138765
12774777120961512082121404sp: PROY_SALTYSalmonella typhimurium proY51.385.0433proline-specific permease
12784778120993412121292196sp: DEAD_KLEPNKlebsiella pneumoniae CG4348.174.3643DEAD box ATP-dependent RNA
DEAD box ATP-dependent RNAhelicase
helicase deaD
1279477912131151212429687prf: 2323363BTMycobacterium leprae24.747.4247bacterial regulatory protein, tetR
B1308_C2_181family
12804780121326912148581590sp: PCPB_FLAS3Sphingomonas flava pcpB24.547.7595pentachlorophenol 4-
monooxygenase
12814781121487112159381068sp: CLCE_PSESBPseudomonas sp. B13 clcE40.472.0354maleylacetate reductase
1282478212159521216836885sp: CATA_ACICAAcinetobacter calcoaceticus30.659.4278catechol 1,2-dioxygenase
catA
1283478312173741216904471
1284478412179821217443540pir: A70672Mycobacterium tuberculosis31.958.4185hypothetical protein
H37Rv Rv2972c
12854785121989512229963102sp: SNF2_YEASTSaccharomyces cerevisiae24.955.4878transcriptional regulator
SNF2
12864786122290512218411065
1287478712229861223843858gp: SCO007731_6Streptomyces coelicolor A3(2)29.656.2203hypothetical protein
orfZ
12884788122388712250591173pir: E70755Mycobacterium tuberculosis39.267.3395phosphoesterase
H37Rv Rv1277
12894789122506612276932628sp: Y084_MYCTUMycobacterium tuberculosis29.759.6915hypothetical protein
H37Rv Rv1278
1290479012275871227282306
1291479112276571227340318
1292479212278631228636774gp: AB029896_1Petroleum-degrading bacterium37.364.6220esterase or lipase
HD-1 hde
1293479312287181229095378
1294479412291501229935786
1295479512297161229180537sp: ATOE_ECOLIStreptomyces coelicolor37.769.7122short-chain fatty acids transporter
SC1C2.14c atoE
1296479612299951230480486sp: PECS_ERWCHErwinia chrysanthemi recS24.756.6166regulatory protein
1297479712306101230831222
1298479812314321230914519
1299479912317301232479750sp: FNR_ECOLIEscherichia coli K12 MG1655 fnr25.057.9228fumarate (and nitrate) reduction
regulatory protein
1300480012326031232836234sp: MERP_SHEPUShewanella putrefaciens merP33.366.781mercuric transort protein periplasmic
component precursor
13014801123300712348811875sp: ATZN_ECOLIEscherichia coli K12 MG165538.070.6605zinc-transporting ATPase Zn(II)-
atzNtranslocating P-type ATPase
1302480212349831235612630sp: RELA_VIBSSVibrio sp. S14 relA32.958.4137GTP pyrophosphokinase (ATP: GTP
3′-pyrophosphotransferase) (ppGpp
synthetase I)
13034803123812512365451581gsp: R80504Streptomyces lividans tap26.649.3601tripeptidyl aminopeptidase
1304480412421561241554603
1305480512422751242156120
1306480612436211243728108GSP: P61449Corynebacterium glutamicum95.098.024homoserine dehydrogenase
13074807124520112439421260
1308480812455321244843690
1309480912464961245720777sp: NARI_BACSUBacillus subtilis narI45.069.6220nitrate reductase gamma chain
1310481012472391246508732sp: NARJ_BACSUBacillus subtilis narJ30.363.4175nitrate reductase delta chain
13114811124879112471991593sp: NARH_BACSUBacillus subtilis narH56.683.4505nitrate reductase beta chain
1312481212498511250444594PIR: D72603Aeropyrum pernix K1 APE129136.048.0137hypothetical protein
1313481312515451251817273PIR: B72603Aeropyrum pernix K1 APE128936.055.083hypothetical protein
13144814125253712487943744sp: NARG_BACSUBacillus subtilis narG46.973.81271nitrate reductase alpha chain
13154815125390612525571350sp: NARK_ECOLIEscherichia coli K12 narK32.867.9461nitrate extrusion protein
1316481612541461254634489sp: CNX1_ARATHArabidopsis thaliana CV cnx132.565.0157molybdopterin biosynthesis cnx1
protein (molybdenum cofactor
biosynthesis enzyme cnx1)
13174817125660212547371866sp: PRTS_SERMASerratia marcescens strain IFO-21.145.9738extracellular serine protease
3046 prtSprecurosor
1318481812570671257750684
13194819125785812568511008sp: Y0D3_MYCTUMycobacterium tuberculosis30.862.6334hypothetical membrane protein
H37Rv Rv1841c
13204820125926512578651401sp: Y0D2_MYCTUMycobacterium tuberculosis31.660.2472hypothetical membrane protein
H37Rv Rv1842c
1321482112599891259429561gp: PPU242952_2Pseudomonas putida mobA27.552.3178molybdopterin guanine dinucleotide
synthase
13224822126120112599931209sp: MOEA_ECOLIMycobacterium tuberculosis32.858.2366molybdoptein biosynthesis protein
H37Rv Rv0438c moeA
13234823126281812616881131sp: CNX2_ARATHArabidopsis thaliana cnx251.473.7354molybdopterin biosynthsisi protein
Moybdenume (mosybdenum
cofastor biosythesis enzyme)
13244824126461012628861725sp: ALKK_PSEOLPseudomonas oleovorans36.765.7572edium-chain fatty acid—CoA ligase
13254825126514212674272286sp: RHO_MICLUMicrococcus luteus rho50.773.8753Rho factor
1326482612656651266267603
1327482712663061265611696
13284828126644912654271023
13294829126743012685031074sp: RF1_ECOLIEscherichia coli K12 RF-141.971.9363peptide chain release factor 1
1330483012685071269343837sp: HEMK_ECOLIEscherichia coli K1231.157.9280protoporphyrinogen oxidase
1331483112690401268267774
1332483212693961270043648sp: YD01_MYCTUMycobacterium tuberculosis62.386.0215hypothetical protein
H37Rv Rv1301
13334833127004712711921146sp: RFE_ECOLIEscherichia coli K12 rfe31.158.4322undecaprenyl-phosphate alpha-N-
acetylglucosaminyltransferase
1334483412712131271698486
1335483512718711272119249GPU: AB046112_1Corynebacterium glutamicum98.099.080hypothetical protein
atpI
1336483612723401273149810sp: ATP6_ECOLIEscherichia coli K12 atpB24.156.7245ATP synthase chain a (protein 6)
1337483712732861273525240sp: ATPL_STRLIStreptomyces lividans atpL54.985.971H+-transporting ATP synthase lipid-
binding protein. ATP synthase C
chane
1338483812735591274122564sp: ATPF_STRLIStreptomyces lividans atpF27.866.9151H+-transporting ATP synthase chain b
1339483912741311274943813sp: ATPD_STRLIStreptomyces lividans atpD34.367.2274H+-transporting ATP synthase delta
chain
13404840127497512766481674sp: ATPA_STRLIStreptomyces lividans atpA66.988.4516H+-transporting ATP synthase alpha
chain
1341484112767081277682975sp: ATPG_STRLIStreptomyces lividans atpG46.376.6320H+-transporting ATP synthase
gamma chain
13424842127768812791361449sp: ATPB_CORGLCorynebacterium glutamicum99.8100.0483H+-transporting ATP synthase beta
AS019 atpBchain
1343484312791511279522372sp: ATPE_STRLIStreptomyces lividans atpE41.073.0122H+-transporting ATP synthase
epsilon chain
1344484412797701280240471sp: Y02W_MYCTUMycobacterium tuberculosis38.667.4132hypothetical protein
H37Rv Rv1312
1345484512802701280959690sp: Y036_MYCTUMycobacterium tuberculosis70.085.7230hypothetical protein
H37Rv Rv1321
1346484612809671281251285GP: SC26G5_35Streptomyces coelicolor A3(2)45.056.095putative ATP/GTP-binding protein
1347484712817141281262453sp: YQJC_BACSUBacillus subtilis yqjC35.868.7134hypothetical protein
1348484812817941282105312sp: YC20_MYCTUMycobacterium tuberculosis54.579.2101hypothetical protein
H37Rv Rv1898
1349484912821941283114921sp: YD24_MYCTUMycobacterium tuberculosis37.971.4301thioredoxin
H37Rv Rv1324
13504850128332412844661143gp: ECO237695_3Escherichia coli K12 ssuD50.374.3366FMNH2-dependent aliphatic
sulfonate monooxygenase
1351485112845171285284768sp: SSUC_ECOLIEscherichia coli K12 ssuC40.875.8240alphatic sulfonates transport
permease protein
1352485212853021286030729sp: SSUB_ECOLIEscherichia coli K12 ssuB50.472.8228alphatic sulfonates transport
permease protein
1353485312860431286999957sp: SSUA_ECOLIEscherichia coli K12 ssuA35.162.1311sulfonate binding protein precursor
13544854128947312872812193sp: GLGB_ECOLIMycobacterium tuberculosis46.172.77101,4-alpha-glucan branching enzyme
H37Rv Rv1326c glgB(glycogen branching enzyme)
13554855129100712895141494sp: AMY3_DICTHDictyoglomus thermophilum22.950.5467alpha-amylase
amyC
1356485612910261291373348
1357485712916991292577879sp: FEPC_ECOLIEscherichia coli K12 fepC31.887.6211ferric enterobactin transport ATP-
binding protein or ABC transport
ATP-binding protein
1358485812932221294025804pir: C70860Mycobacterium tuberculosis39.668.5260hypothetical protein
H37Rv Rv3040c
13594859129415112952061056pir: H70859Mycobacterium tuberculosis43.170.0367hypothetical protein
H37Rv Rv3037c
1360486012950471294436612
1361486112954351296220786sp: FIXA_RHIMERhizobium meliloti fixA31.264.8244electron transfer flavoprotein beta-
subunit
1362486212962531297203951sp: FIXB_RHIMERhizobium meliloti fixB33.161.8335electron transfer flavoprotein alpha
subunit for various dehydrogenases
1363486312964791297093615
13644864129721212983391128sp: NIFS_AZOVIAzotobacter vinelandii nifS35.267.7375nitrogenase cofactor sythesis protein
1365486512986531298342312
13664866130014512990001146sp: Y4ME_RHISNRhizobium sp. NGR234 plasmid29.555.7397hypothetical protein
pNGR234a y4mE
1367486713003691300145225sp: Y4MF_RHISNRhizobium sp. NGR234 plasmid47.576.359transcriptional regulator
pNGR234a Y4mF
1368486813005521301055504sp: YHBS_ECOLIEscherichia coli K12 MG165534.855.3181acetyltransferase
1369486913019291300988942
13704870130312313019751149
1371487113032991303694396
13724872130382913049231095pir: C70858Mycobacterium tuberculosis61.880.9361tRNA (5-methylaminomethyl-2-
H37Rv Rv3024cthiouridylate)-methyltransferase
1373487313045361303883654
1374487413049321305921990pir: B70857Mycobacterium tuberculosis33.766.0332hypothetical protein
H37Rv Rv3015c
13754875130738413059241461sp: TCMA_STRGAStreptomyces glaucescens tcmA30.265.8500tetracenomycin C resistance and
export protin
1376487613081961307462735
13774877130833013103692040sp: DNLJ_RHOMRRhodothermus marinus dnlJ42.870.6677DNA ligase
(polydeoxyribonucleotide synthase
[NAD+]
1378487813110971310435663pir: H70856Mycobacterium tuberculosis40.070.9220hypothetical protein
H37Rv Rv3013
1379487913113201311616297sp: GATC_STRCOStreptomyces coelicolor A3(2)53.064.097glutamyl-tRNA(Gln)
gatCamidotransferase subunit C
13804880131162513131151491sp: GATA_MYCTUMycobacterium tuberculosis74.083.0484glutamyl-tRNA(Gln)
H37Rv gatAamidotransferase subunit A
1381488113132701314118849sp: VIUB_VIBVUVibrio vulnificus viuB28.154.0263vibriobactin utilization protein/iron-
chelator utilization protein
1382488213147751314470306gp: SCE6_24Streptomyces coelicolor A3(2)46.979.296hypothetical membrane protein
SCE6.24
13834883131501313160831071sp: PFP_AMYMEAmycolatopsis methanolica pfp54.877.9358pyrophosphate—fructose 6-
phosphate 1-phosphotransrefase
1384488413159541315325630
13854885131633813174441107sp: CCPA_BACMEBacillus megaterium ccpA31.431.4328glucose-resistance amylase
regulator (catabolite control protein)
13864886131743413190051572sp: RBSA_ECOLIEscherichia coli K12 rbsA44.776.2499ripose transport ATP-binding protein
1387488713190051319976972sp: RBSC_ECOLIEscherichia coli K12 MG165545.676.9329high affinity ribose transport protein
rbsC
1388488813200011320942942sp: RBSB_ECOLIEscherichia coli K12 MG165545.977.7305periplasmic ribose-binding protein
rbsB
1389488913209521321320369sp: RBSD_ECOLIEscherichia coli K12 MG165541.768.4139high affinity ribose transport protein
rbsD
1390489013214761322111636sp: YIW2_YEASTSaccharomyces cerevisiae31.058.0200hypothetical protein
YIR042c
13914891132239313234061014gp: SCF34_13Streptomyces coelicolor31.460.2354iron-siderophore binding lipoprotein
SCF34.13c
13924892132353313245371005sp: NTCI_RATRattus norvegicus (Rat) NTCI35.861.9268Na-dependent blle acid transporter
13934893132477813262561479gsp: W61467Staphylococcus aureus WHU 2943.171.8485RNA-dependent amidotransferase B
ratB
1394489413263781327049672sp: F4RE_METJAMethanococcus jannaschii 32.661.1172putative F420-dependent NADH
MJ1501 f4rereductase
13954895133096713298911077sp: YQJG_ECOLIEscherichia coli K12 yqjG39.866.9317hypothetical protein
1396489613311021331875774pir: A70672Mycobacterium tuberculosis39.362.4234hypothetical protein
H37Rv Rv2972c
13974897133195313330081056pir: H70855Mycobacterium tuberculosis27.452.6325hypothetical membrane protein
H37Rv Rv3005c
1398489813334241333188237
13994899133528013334421839gp: AJ012293_1Corynebacterium glutamicum99.299.4613dihydroxy-acid dehydratase
ATCC 13032 llvD
1400490013359751335412564pir: G70855Mycobacterium tuberculosis33.368.6105hypothetical protein
H37Rv Rv3004
14014901133756713360951473sp: YILV_CORGLCorynebacterium glutamicum100.0100.062hypothetical membrane protein
ATCC 13032 yilV
1402490213386091338379231GP: SSU18930_263Sulfolobus solfataricus45.055.066hypothetical protein
1403490313420721342677606
1404490413424571341960498sp: NRTD_SYNP7Synechococcus sp. nrtD50.980.8167nitrate transport ATP-binding potein
1405490513427271342461267sp: MALK_ENTAEEnterobacter aerogenes46.078.287maltose/maltodextrin transport ATP-
(Aerobacter aerogenes) malKbinding protein
1406490613436751342794882sp: NRTA_ANASPAnabaena sp. strain PCC 712028.156.8324nitrate transporter protein
nrtA
1407490713440181344464447
1408490813444401344808369
1409490913449351345420486sp: DIM6_STRCOStreptomyces coelicolor39.473.2142actinorhodin polyketide dimerase
1410491013454861346439954sp: CZCD_ALCEURalstonia eutropha czcD39.172.7304cobalt-zinc-cadimium resistance
protein
1411491113454871345335153
1412491213463311345642690
14134913134645813482721815sp: V686_METJAMethanococcus jannaschii 22.953.7642hypothetical protein
14144914134833413500761743
14154915135085513524441590gsp: Y22646Brevibacterium flavum serA99.8100.0530D-3-phosphoglycerate
dehydrogenase
1416491613520531351727327SP: YEN1_SCHPOSchizosaccharomyces pombe29.052.0105hypothetical serine-rich protein
SPAC11G7.01
1417491713525851353451867
14184918135560113545401062
14194919135568913575541866pir: T03476Rhodobacter capsulatus strain32.963.1620hypothetical protein
SB1003
1420492013564521356853402
1421492113575571358210654
1422492213582591359062804sp: HPCE_ECOLIEscherichia coli C hpcE33.359.2228homoprotocatechiuate catabolism
bifunctional
isomerase/decarboxylase [includes:
2-hydroxyhepta-2,4-diene-1,7-dioate
isomerase(hhdd isomerase); 5-
carboxymethyl-2-oxo-hex-3-ene-1,7-
dioate decarboxylase(opet
decarboxylase)]
1423492313590521359669618sp: UBIG_ECOLIEscherichia coli K1223.455.7192methyltransferase or 3-
demethylubiquinone-9 3-O-
methyltransferase
14244924136129513601681128sp: DHBC_BACSUBacillus subtilis dhbC38.070.4371isochorismate synthase
14254925136136113628481488sp: SYE_BACSUBacillus subtilis gltX37.369.7485glutamyl-tRNA synthetase
1426492613631381362926213gp: SCJ33_10Streptomyces coelicolor A3(2)77.090.067transcriptional regulator
1427492713636571363142516
1428492813642531363732522
1429492913649151365256342
1430493013649601364340621
1431493113651801364878303
1432493213653961365217180
1433493313658081366137330
1434493413672931367505213
1435493513680701367888183
1436493613680781368395318
14374937136840013695511152
1438493813695511369874324
14394939137163713698771761sp: THIC_BACSUBacillus subtilis thiA or thiC65.181.0599thiamin biosynthesis protein
1440494013723261371979348
1441494113726011373131531
1442494213737981373929132GSP: Y37857Chlamydia trachomatis61.074.044lipoprotein
1443494313745561375491936
14444944137577613733502427sp: PHS1_RATRattus norvegicus (Rat)44.274.0797glycogen phosphorylase
1445494513759871375805183
1446494613760881375933156
14474947137755513761491407sp: YRKH_BACSUBacillus subtilis yrkH25.452.8299hypothetical protein
1448494813784151377666750sp: Y441_METJAMethanococcus jannaschii Y44125.464.8256hypothetical membrane protein
1449494913789421378466477
1450495013790031379566564sp: SPOT_ECOLIEscherichia coli K12 spoT29.860.1178guanosine 3′,5′-bis(diphosphate) 3′-
pyrophosphatase
1451495113802591379555705sp: ICLR_ECOLIEscherichia coli K12 iclR26.160.7257acetate repressor protein
14524952138044013818821443sp: LEU2_ACTTIActinoplanes teichomyceticus68.187.54733-isopropylmalate dehydratase large
leu2subunit
1453495313819021382492591sp: LEUD_SALTYSalmonella typhimurium67.789.21953-isopropylmalate dehydratase small
subunit
1454495413828191382502318
1455495513837981382845954gp: MLCB637_35Mycobacterium tuberculosis45.971.4294mutator mutT protein ((7,8-dihydro-
H37Rv MLCB637.35c8-oxoguanine-triphosphatase)(8-
oxo-dGTPase)(dGTP
pyrophosphohydrolase)
1456495613839301384085156
1457495713841301385125996sp: GPDA_BACSUBacillus subtilis gpdA45.072.2331NAD(P)H-dependent
dihydroxyacetone phosphate
reductase
14584958138515313862321080sp: DDLA_ECOLIEscherichia coli K12 MG165540.467.4374D-alanine-D-alanine ligase
ddlA
1459495913872701386293978
1460496013873321388324993sp: THIL_ECOLIEscherichia coli K12 thiL32.257.6335thiamin-phosphate kinase
1461496113883121389073762sp: UNG_MOUSEMus musculus ung38.859.6245uracil-DNA glycosylase precursor
14624962138920813907881581sp: Y369_MYCGEMycoplasma genitalium (SGC3)23.156.3568hypothetical protein
MG369
14634963139079613929162121sp: RECG_ECOLIEscherichia coli K12 recG35.460.0693ATP-dependent DNA helicase
1464496413919611391638324GSP: Y75303Neisseria meningitidis31.048.0108polypeptides predicted to be useful
antigens for vaccines and
diagnostics
1465496513929391393151213sp: BCCP_PROFRPropionibacterium freudenreichii38.867.267biotin carboxyl carrier protein
subsp. Shermanii
1466496613931541393735582sp: YHHF_ECOLIEscherichia coli K12 yhhF37.163.5167methylase
1467496713937421394221480sp: KDTB_ECOLIEscherichia coli K12 MG165542.678.7155lipopolysaccharide core biosynthesis
kdtBprotein
14684968139485413959331080
1469496913948941395097204GSP: Y75358Neisseria gonorrhoeae67.074.065Neisserial polypeptides predicted to
be useful antigens for vaccines and
diagnostics
1470497013955491394800750sp: GLNQ_BACSTBacillus stearothermophilus56.478.6252ABC transporter or glutamine ABC
glnQtransporter, ATP-binding protein
1471497113964101395568843sp: NOCM_AGRT5Agrobacterium tumefaciens32.775.0220nopaline transport protein
nocM
1472497213974211396561861sp: GLNH_ECOLIEscherichia coli K12 MG165527.459.0234glutamine-binding protein precursor
glnH
1473497313976621398468807
1474497413995341398557978pir: H69160Methanobacterium28.660.3322hypothetical membrane protein
thermoautotrophicum MTH465
1475497514009261401333408
1476497614009401400185756sp: VINT_BPL54Bacteriophage L54a vinT26.952.5223phage integrase
1477497714013331402076744
1478497814022721402703432
1479497914028741402368507
1480498014031281403991864
1481498114039971404215219
1482498214048851404694192pir: S60890Corynebacterium glutamicum88.596.226insertion element (IS3 related)
orf2
1483498314061741405320855
1484498414071091406999111PIR: S60890Corynebacterium glutamicum89.097.037hypothetical protein
1485498514075351407167369
1486498614078731407559315
1487498714090231408703321
1488498814098021409428375
1489498914110111410064948
1490499014114241411119306
1491499114120001411437564
1492499214123511412572222
1493499314129161412626291
14944994141374514164592715sp: DPO1_MYCTUMycobacterium tuberculosis56.380.8896DNA polymerase I
polA
14954995141788314164621422sp: CMCT_NOCLAStreptomyces lactamdurans33.867.8456cephamycin export protein
cmcT
1496499614179621418870909gp: SCJ9A_15Streptomyces coelicolor A3(2)41.365.4283DNA-binding protein
SCJ9A.15c
1497499714188761419748873sp: MORA_PSEPUPseudomonas putida morA46.576.1284morphine-6-dehydrogenase
1498499814200361419878159
1499499914207241420071654sp: YAFE_ECOLIStreptomyces coelicolor31.958.3163hypothetical protein
SCH5.13 yafE
15005000142109914225561458sp: RS1_ECOLIEscherichia coli K12 rpsA39.571.445130S ribosomal protein S1
15015001142257114210961476
1502500214252791425878600sp: YACE_BRELABrevibacterium lactofermentum80.593.9195hypothetical protein
ATCC 13869 yacE
15035003142625714273541098
1504500414279571427376582
1505500514280491427804246
1506500614282901429246957
1507500714291591428224936sp: IUNH_CRIFACrithidia fasciculata iunH61.981.0310inosine-uridine preferring nucleoside
hypolase (purine nucleosidase)
15085008143064214291941449sp: QACA_STAAUStaphylococcus aureus 23.653.8517aniseptic resistance protein
1509500914315791430659921sp: RBSK_ECOLIEscherichia coli K12 rbsK35.567.6293ribose kinase
15105010143261214315751038sp: ASCG_ECOLIEscherichia coli K12 ascG30.065.6337criptic asc operon repressor,
ranscription regulator
1511501114327501433547798
15125012143410514362012097sp: UVRB_STRPNStreptococcus pneumoniae57.483.3671excinuclease ABC subunit B
plasmid pSB470 uvrB
1513501314363351436775441sp: Y531_METJAMethanococcus jannaschii33.659.2152hypothetical protein
MJ0531
1514501414372491436869381sp: YTFH_ECOLIEscherichia coli K12 ytfH38.880.2121hypothetical protein
1515501514373561438201846sp: YTFG_ECOLIEscherichia coli K12 ytfG53.877.1279hypothetical protein
1516501614393431440026684
15175017144056014382122349pir: H70040Bacillus subtilis yvgS23.247.2839hypothetical protein
1518501814415861440675912gp: SC9H11_26Streptomyces coelicolor A3(2)32.768.0150hypothetical protein
SC9H11.26c
1519501914423921441793600sp: YCBL_ECOLIEscherichia coli K12 ycbL30.458.4214hydrolase
15205020144248714453332847sp: UVRA_ECOLIEscherichia coli K12 uvrA56.280.6952excinuclease ABC subunit A
1521502114441151443810306PIR: JQ0406Micrococcus luteus 40.057.0100hypothetical protein 1246 (uvrA
region)
1522502214453931444944450PIR: JQ0406Micrococcus luteus 31.047.0142hypothetical protein 1246 (uvrA
region)
1523502314461581446874717
15245024144744614453232124
1525502514477921448358567sp: IF3_RHOSHRhodobacter sphaeroides infC52.578.2179translation initiation factor IF-3
1526502614483901448581192sp: RL35_MYCFEMycoplasma fermentans41.776.76050S ribosomal protein L35
1527502714486451449025381sp: RL20_PSESYPseudomonas syringae pv.75.092.711750S ribosomal protein L20
syringae
1528502814499401449119822
1529502914501261450692567
1530503014509181451820903sp: UGPA_ECOLIEscherichia coli K12 MG165533.271.6292sn-glycerol-3-phosphate transport
ugpAsystem permease protein
1531503114518201452653834sp: UGPE_ECOLIEscherichia coli K12 MG165533.370.4270sn-glycerol-3-phosphate transport
upgEsystem protein
15325032145275814540711314sp: UGPB_ECOLIEscherichia coli K12 MG165526.657.6436sn-glycerol-3-phosphate transport
ugpBsystem permease proein
15335033145411514553381224sp: UGPC_ECOLIEscherichia coli K12 MG165544.071.3393sn-glycerol-3-phosphate transport
ugpCATP-binding protein
1534503414543501454102249PIR: E72756Aeropyrum pernix K1 APE004247.056.074hypothetical protein
1535503514560661455350717sp: GLPQ_BACSUBacillus subtilis glpQ26.250.0244glycerophosphoryl diester
phosphodiesterase
1536503614563551456948594sp: TRMH_ECOLIEscherichia coli K12 MG165534.071.2153tRNA(guanosine-2′-0-)-
trmHmethlytransferase
15375037145704714580661020sp: SYFA_BACSUBacillus subtilis 168 syfAphenylalanyl-tRNA synthetase alpha
chain
15385038145813314606162484sp: SYFB_ECOLIEscherichia coli K12 MG165542.671.7343phenylalanyl-tRNA synthetase beta
syfBchain
1539503914589661458196771
1540504014611571462128972sp: ESTA_STRSCStreptomyces scabies estA26.555.1363esterase
15415041146213414635161383sp: MDMB_STRMYStreptomyces mycarofaciens30.056.3423macrolide 3-O-acyltransferase
mdmB
1542504214635331463934402
15435043146408314651231041gp: AF005242_1Corynebacterium glutamicum98.399.1347N-acetylglutamate-5-semialdehyde
ASO19 argCdehydrogenase
15445044146521014663731164sp: ARGJ_CORGLCorynebacterium glutamicum99.599.7388glutamate N-acetyltransferase
ATCC 13032 argJ
15455045146737614685481173sp: ARGD_CORGLCorynebacterium glutamicum99.099.2391acetylornithine aminotransferase
ATCC 13032 argD
15465046147021114714131203sp: ASSY_CORGLCorynebacterium glutamicum99.599.5401argininosuccinate synthetase
ASO19 argG
15475047147136214701541209
15485048147147714729071431gp: AF048764_1Corynebacterium glutamicum83.390.0478argininosuccinate lyase
ASO19 argH
15495049147297714741191143
15505050147411914756931575
1551505114756831476294612
1552505214763431476519177sp: YCAR_ECOLIEscherichia coli K12 ycaR48.072.050hypothetical protein
15535053147655014778091260sp: SYY1_BACSUBacillus subtilis syy148.479.6417tyrosyl-tRNA synthase (tyrosine—
tRNA ligase)
1554505414783931477929465sp: Y531_METJAMethanococcus jannaschii26.964.4149hypothetical protein
MJ0531
1555505514788921478503390
1556505614834751483335141PIR: F81737Chlamydia muridarum Nigg71.075.042hypothetical protein
TC0129
1557505714839961483724273GSP: Y35814Chlamydia pneumoniae61.066.084hypothetical protein
15585058148467514860271353sp: IF2_BORBUBorrelia burgdorferi IF236.367.0182translation initiation factor IF-2
1559505914860421487025984sp: YZGD_BACSUBacillus subtilis yzgD29.660.1311hypothetical protein
1560506014870321487193162
1561506114872381488056819sp: YQXC_BACSUBacillus subtilis yqxC38.569.6260hypothetical protein
1562506214881461489018873sp: YFJB_HAEINMycobacterium tuberculosis31.631.6225hypothetical protein
H37Rv Rv1695
15635063148910314908811779sp: RECN_ECOLIEscherichia coli K12 recN31.463.4574DNA repair protein
15645064149094414921341191pir: H70502Mycobacterium tuberculosis41.973.1394hypothetical protein
H37Rv Rv1697
1565506514921471493109963pir: A70503Mycobacterium tuberculosis30.468.1313hypothetical protein
H37Rv Rv1698
15665066149351314951741662sp: PYRG_ECOLIEscherichia coli K12 pyrG55.076.7549CTP synthase (UTP—ammonia
ligase)
1567506714952051495861657sp: YQKG_BACSUBacillus subtilis yqkG36.371.3157hypothetical protein
1568506814958611496772912gp: AF093548_1Staphylococcus aureus xerD39.771.7300tyrosine recombinase
15695069149832414967951530sp: TLRC_STRFRStreptomyces fradiae tlrC30.559.7551tyrosin resistance ATP-binding
protein
1570507014988631499645783gp: CCU87804_4Caulobacter crescentus parA44.673.6258chromosome partitioning protein or
ATPase involved in active
partitioning of diverse bacterial
plasmids
1571507114999311500695765sp: YPUG_BACSUBacillus subtilis ypuG28.364.5251hypothetical protein
1572507215014711500911561
1573507315017101502576867gp: AF109156_1Datisca glomerata tst35.667.0270thiosulfate sulfurtransferase
1574507415026341503176543sp: YPUH_BACSUBacillus subtilis ypuH33.165.7172hypothetical protein
1575507515034831504238756sp: RLUB_BACSUBacillus subtilis rluB45.972.5229ribosomal large subunit
pseudouridine synthase B
1576507615042561504945690sp: KCY_BACSUBacillus subtilis cmk38.673.6220cytidylate kinase
15775077150501715065731557sp: YPHC_BACSUBacillus subtilis yphC42.874.0435GTP binding protein
1578507815073271506662666
1579507915079021507405498
1580508015087291507917813sp: YX42_MYCTUMycobacterium tuberculosis36.267.2232methyltransferase
Rv3342
15815081150881315103661554prf: 2513302BCorynebacterium striatum M82B29.760.1499ABC transporter
tetA
15825082151036615121321767prf: 2513302ACorynebacterium striatum M82B31.256.3602ABC transporter
tetB
1583508315116671510843825
1584508415121891512977789sp: YGIE_ECOLIEscherichia coli K12 ygiE39.773.2257hypothetical membrane protein
1585508515145051514693189
15865086151452715129801548gp: AB029555_1Bacillus subtilis ATCC 937225.761.5499Na+/H+ antiporter
nhaG
1587508715151591514974186
1588508815153961515815420
1589508915157821515408375sp: YCHJ_ECOLIEscherichia coli K12 o249#936.957.7130hypothetical protein
ychJ
15905090151696215157991164pir: C69334Archaeoglobus fulgidus AF067525.263.82102-hydroxy-6-oxohepta-2,4-dienoate
hydrolase
15915091151717015194582289sp: SECA_BACSUBacillus subtills secA35.261.7805preprotein translocase SecA subunit
1592509215196011520029429gp: AF173844_2Mycobacterium smegmatis garA75.893.2132signal transduction protein
1593509315201901520945756sp: Y0DF_MYCTUMycobacterium tuberculosis41.974.4234hypothetical protein
H37Rv Rv1828
1594509415209571521589633sp: Y0DE_MYCTUMycobacterium tuberculosis30.863.2133hypothetical protein
H37Rv Rv1828
1595509515217711522343573sp: Y0DE_MYCTUMycobacterium tuberculosis71.484.3178hypothetical protein
H37Rv Rv1828
1596509615229411522432510
15975097152450015230521449
1598509815253741525973600
1599509915254971524568930
16005100152653415254731062sp: YHDP_BACSUBacillus subtilis yhdP33.969.0342hemolysin
16015101152791315265341380sp: YHDT_BACSUBacillus subtilis yhdT31.465.565hemolysin
1602510215279681528186219
16035103152933015279871344gp: TTHERAGEN_1Thermus thermophilus herA41.269.5374DEAD box RNA helicase
1604510415294861530220735sp: YD48_MYCTUMycobacterium tuberculosis34.366.1245ABC transporter ATP-binding protein
H37Rv Rv1348
16055105153181615303411476gsp: W27613Brevibacterium flavum99.099.24926-phosphogluconate dehydrogenase
1606510615319331532394462pir: G70664Mycobacterium tuberculosis39.767.8121thioesterase
H37Rv Rv1847
1607510715323221532996675
1608510815330411533781741sp: NODI_RHIS3Rhizobium sp. N33 nodl39.668.1235nodulation ATP-binding protein I
1609510915337811534521741pir: E70501Mycobacterium tuberculosis43.176.3232hypothetical membrane protein
H37Rv Rv1686c
1610511015354011534529873sp: YFHH_ECOLIEscherichia coli K12 yfhH26.763.9277transcriptional regulator
1611511115362271535382846sp: PHNE_ECOLIEscherichia coli K12 phnE29.963.4281phosphonates transport system
permease protein
1612511215370301536227804sp: PHNE_ECOLIEscherichia coli K12 phnE27.262.3268phosphonates transport system
permease protein
1613511315378331537030804sp: PHNC_ECOLIEscherichia coli K12 phnC44.872.0250phosphonates transport ATP-binding
protein
1614511415387591538968210
16155115153891915378701050
1616511615396641538963702
16175117154140315398201584sp: THID_SALTYSalmonella typhimurium thiD47.370.2262phosphomethylpyrimidine kinase
1618511815429221542119804sp: THIM_SALTYSalmonella typhimurium LT246.677.5249hydoxyethylthiazole kinase
thiM
16195119154497615462891314pir: H70830Mycobacterium tuberculosis28.655.0451cyclopropane-fatty-acyl-phospholipid
H37Rv ufaA1synthase
16205120154769215463071386prf: 2223339BBurkholderia cepacia Pc70132.566.9468sugar transporter or 4-methyl-o-
mopBphthalate/phthalate permease
1621512115484401547967474prf: 2120352BThermus flavus AT-62 gpt36.559.0156purine phosphoribosyltransferase
1622512215486511549349699sp: YEBN_ECOLIEscherichia coli K12 yebN39.868.5206hypothetical protein
1623512315494031550398996gp: AF178758_2Sinorhizobium sp. As4 arsB23.354.6361arsenic oxyanion-translocation pump
membrane subunit
1624512415504691550951483
1625512515515451552237693gp: SCI7_33Streptomyces coelicolor A3(2)62.283.8222hypothetical protein
SCI7.33
16265126155251815539721455gp: PSTRTETC1_6Pseudomonas sp. R9 ORFA51.883.6469sulfate permease
1627512715537221553297426GP: PSTRTETC1_7Pseudomonas sp. R9 ORFG39.050.097hypothetical protein
1628512815546841554070615
1629512915548611555067207
1630513015550791554891189
1631513115558351555086750
1632513215563761556771396pir: A70945Mycobacterium tuberculosis71.887.3110hypothetical protein
H37Rv Rv2050
1633513315578231557014810prf: 2317468ASchizosaccharomyces pombe39.271.0217dolichol phosphate mannose
dpm1synthase
16345134155949315578591635sp: LNT_ECOLIEscherichia coli K12 Int25.155.6527apolipoprotein N-acyltransferase
1635513515602371559497741
16365136156166015604371224gp: AF188894_1Candida albicans lip123.755.6392secretory lipase
1637513715617801562553774pir: C70764Mycobacterium tuberculosis31.356.7291precorrin 2 methyltransferase
H37Rv cobG
16385138156380215625251278sp: COBL_PSEDEPseudomonas denitrificans32.460.8411precorrin-6Y C5, 15-
SC510 cobLmethyltransferase
1639513915638721564237366
1640514015642371564482246
1641514115653021564565738sp: YY12_MYCTUMycobacterium tuberculosis54.175.4244oxidoreductase
H37Rv RV3412
16425142156643815653021137gp: AF014460_1Streptococcus mutans LT1136.161.3382dipeptidase or X-Pro dipeptidase
pepQ
1643514315664681567106639
16445144156990315671172787sp: MTR4_YEASTSaccharomyces cerevisiae26.555.71030ATP-dependent RNA helicase
YJL050W dob1
16455145157093315699321002sp: TATC_ECOLIEscherichia coli K12 tatC28.762.7268sec-independent protein translocase
protein
1646514615713821571068315sp: YY34_MYCLEMycobacterium leprae44.769.485hypothetical protein
MLCB2533.27
1647514715724861571506981sp: YY35_MYCTUMycobacterium tuberculosis31.961.2317hypothetical protein
H37Rv Rv2095c
1648514815734631572492972sp: YY36_MYCLEMycobacterium leprae32.464.8324hypothetical protein
MLCB2533.25
16495149157491515734911425sp: YY37_MYCTUMycobacterium tuberculosis53.177.3467hypothetical protein
H37Rv Rv2097c
1650515015749571575205249
1651515115751361574945192pir: B70512Mycobacterium tuberculosis54.180.361hypothetical protein
H37Rv Rv2111c
16525152157694715754061542pir: C70512Mycobacterium tuberculosis48.674.2516hypothetical protein
H37Rv Rv2112c
1653515315773271577806480PIR: H72504Aeropyrum pernix K1 APE201442.050.0159hypothetical protein
16545154157853115769511581prf: 2422382QRhodococcus erythropolis arc51.678.5545AAA family ATPase (chaperone-like
function)
1655515515794001578567834pir: S72844Mycobacterium leprae pimT57.379.0281protein-beta-aspartate
methyltransferase
16565156158077115794491323gp: AF005050_1Homo sapiens38.167.2436aspartyl aminopeptidase
1657515715808071581640834pir: B70513Mycobacterium tuberculosis45.471.4269hypothetical protein
H37Rv Rv2119
1658515815818511582114264sp: VAPI_BACNODichelobacter nodosus A19840.672.569virulence-associated protein
vapI
16595159158348115822731209prf: 2513299AStaphylococcus aureus norA2321.861.0385quinolon resistance protein
16605160158549015839131578sp: ASPA_CORGLCorynebacterium glutamicum99.899.8526aspartate ammonia-lyase
(Brevibacterium flavum) MJ233
aspA
1661516115864451585603843gp: AF050166_1Corynebacterium glutamicum96.897.5281ATP phosphoribosyltransferase
ASO19 hisG
1662516215875041586812693pir: H72277Thermotoga maritima MSB830.863.1195beta-phosphoglucomutase
TM1254
16635163159123515875733663sp: METH_ECOLIEscherichia coli K12 metH31.662.412545-methyltetrahydrofolate—
homocysteine methyltransferase
1664516415913431591912570
16655165159296615919411026sp: AHPF_XANCHXanthomonas campestris ahpF22.449.5366alkyl hydroperoxide reductase
subunit F
16665166159333715945121176sp: ACR3_YEASTSaccharomyces cerevisiae33.063.9388arsenical-resistance protein
S288C YPR201W acr3
1667516715945321594951420sp: ARSC_STAAUStaphylococcus aureus plasmid32.664.3129arsenate reductase
pl258 arsC
1668516815950301595668639pir: G70964Mycobacterium tuberculosis47.275.6123arsenate reductase
H37Rv arsC
1669516915962211595844378
16705170159746015962491212sp: SYC_ECOLIEscherichia coli K12 cysS35.964.3387cysteinyl-tRNA synthetase
1671517115986231597745879sp: BACA_ECOLIEscherichia coil K12 bacA37.369.4255bacitracin resistance protein
1672517215986671599614948prf: 2214302FAgrobacterium tumefaciens33.462.6326oxidoreductase
mocA
1673517315996791600677999pir: F70577Mycobacterium tuberculosis27.053.5359lipoprotein
H37Rv lppL
16745174160069216018041113sp: PYRD_AGRAEAgrocybe aegerita ura144.067.1334dihydroorotate dehydrogenase
1675517516022811601931351
1676517616026601603466807
16775177160352016046291110gp: PSESTBCBAD_1Pseudomonas syringae tnpA34.755.3360transposase
1678517816053151604830486
1679517916058111605281531sp: YBHB_ECOLIEscherichia coli K12 ybhB44.175.0152bio operon ORF I (biotin biosynthetic
enzyme)
1680518016059611606689729GSP: Y74829Neisseria meningitidis26.033.0198Neisserial polypeptides predicted to
be useful antigens for vaccines and
diagnostics
1681518116076461608248603
16825182160765716058611797prf: 2513302ACorynebacterium striatum M82B43.668.7597ABC transporter
tetB
1683518316090871609335249
16845184160924716076611587prf: 2513302BCorynebacterium striatum M82B36.867.1535ABC transporter
tetA
1685518516101921609842351
1686518616102361610844609pir: JU0052Streptomyces anulatus pac32.456.456puromycin N-acetyltransferase
16875187161223816111501089sp: ARGK_ECOLIEscherichia coli K12 argK43.172.3339LAO(lysine, arginine, and
ornithine)/AO (arginine and
ornithine)transport system kinase
16885188161444416122342211sp: MUTB_STRCMStreptomyces cinnamonensis72.287.5741methylmalonyl-CoA mutase alpha
A3823.5 mutBsubunit
16895189161629816144511848sp: MUTA_STRCMStreptomyces cinnamonensis41.668.2610methylmalonyl-CoA mutase beta
A3823.5 mutAsubunit
1690519016165781617300723sp: YS13_MYCTUMycobacterium tuberculosis39.770.1224hypothetical membrane protein
H37Rv Rv1491c
1691519116173981617994597
16925192161961616183211296sp: YS09_MYCTUMycobacterium tuberculosis64.187.0370hypothetical membrane protein
H37Rv Rv1488
1693519316201061619672435pir: B70711Mycobacterium tuberculosis44.778.7141hypothetical membrane protein
H37Rv Rv1487
1694519416210091620167843gp: SCC77_24Streptomyces coelicolor A3(2)51.072.8261hypothetical protein
SCC77.24
1695519516210561621838783
16965196162295016218411110sp: HEMZ_PROFRPropionibacterium freudenreichii36.865.7364ferrochelatase
subsp. Shermanii hemH
16975197162482616230271800sp: P54_ENTFCStreptococcus faecium25.556.5611invasin
1698519816259251625428498
16995199162627916291072829pir: F70873Mycobacterium tuberculosis69.985.9959aconitate hydratase
H37Rv acn
1700520016292981629861564pir: E70873Mycobacterium tuberculosis54.681.6174transcriptional regulator
H37Rv Rv1474c
1701520116299131630668756pir: F64496Methanococcus jannaschii21.351.9235GMP synthetase
MJ1575 guaA
1702520216313291630667663gp: SCD82_4Streptomyces coelicolor A3(2)32.662.0221hypothetical protein
SCD82.04c
1703520316316601631926267pir: E64494Methanococcus jannaschii37.280.286hypothetical protein
MJ1558
1704520416317451631353393
17055205163193316333241392gp: AE002515_9Neisseria meningitidis MC5861.286.1446hypothetical protein
NMB1652
1706520616325881632109480GSP: Y38838Neisseria gonorrhoeae ORF2454.060.0113antigenic protein
1707520716331371632682456GSP: Y38838Neisseria gonorrhoeae59.069.0152antigenic protein
17085208163356616362412676sp: ATA1_SYNY3Synechocystis sp. PCC680342.673.2883cation-transporting ATPase P
sll1614 pma1
1709520916345631633781783
1710521016367321636244489gp: SC3D11_2Streptomyces coelicolor A3(2)35.858.3120hypothetical protein
SC3D11.02c
17115211163708116384421362
1712521216391321638776357
1713521316393651639520156
1714521416396561639817162
1715521516397811640155375prf: 2408488HStreptococcus thermophilus43.073.8107host cell surface-exposed lipoprotein
phage TP-J34
1716521616405461641001456prf: 2510491ACorynephage 304L int34.460.4154integrase
17175217164267416410461629sp: YJJK_ECOLIEscherichia coli K12 yjjK32.864.4497ABC transporter ATP-binding protein
17185218164421816427431476
17195219164549916443181182sp: NANH_MICVIMicromonospora viridifaciens51.972.4387sialidase
ATCC 31146 nedA
1720522016456611646368708gp: AF121000_8Corynebacterium glutamicum99.6100.0236transposase (IS1628)
22243 R-plasmid pAG1 tnpB
1721522116458211646063243GPU: AF164956_23Corynebacterium glutamicum64.072.037transposase protein fragment
TnpNC
1722522216458611645601261GP: NT1TNIS_5Plasmid NTP1632.043.088hypothetical protein
1723522316465491647133585
1724522416476341647212423pir: B75015Pyrococcus abyssi Orsay32.770.1107dTDP-4-keto-L-rhamnose reductase
PAB1087
1725522516480971647651447pir: S72754Mycobacterium leprae63.885.2149nitrogen fixation protein
MLCL536.24c nifU7
1726522616485481648709162PIR: C72506Aeropyrum pernix K1 APE202548.057.052hypothetical protein
17275227164936216481001263pir: S72761Mycobacterium leprae nifS64.784.4411nitrogen fixation protein
1728522816501221649367756gp: SCC22_4Streptomyces coelicolor A3(2)70.289.3252ABC transporter ATP-binding protein
SCC22.04c
17295229165142416502491176pir: A70872Mycobacterium tuberculosis55.283.0377hypothetical protein
H37Rv Rv1462
17305230165287516514331443sp: Y074_SYNY3Synechocystis sp. PCC680341.073.0493ABC transporter
slr0074
1731523116535861652894693gp: SCC22_8Streptomyces coelicolor A3(2)46.171.4217DNA-binding protein
SCC22.08c
17325232165404316556711629pir: F70871Mycobacterium tuberculosis36.367.8518hypothetical membrane protein
H37Rv Rv1459c
17335233165568116567001020pir: S72783Mycobacterium leprae50.277.3317ABC transporter
MLCL536.31 abc2
1734523416567121657515804pir: S72778Mycobacterium leprae41.074.8266hypothetical protein
MLCL536.32
1735523516576771658675999pir: C70871Mycobacterium tuberculosis43.074.6291hypothetical protein
H37Rv Rv1456c
1736523616594961659140357
17375237165950816611361629pir: C71156Pyrococcus horikoshii PH045023.451.0418helicase
1738523816615781662552975sp: QOR_ECOLIEscherichia coli K12 qor37.570.9323quinone oxidoreductase
1739523916635981662630969gp: NWCOXABC_3Nitrobacter winogradskyi coxC37.666.8295cytochrome o ubiquinol oxidase
assembly factor/heme O
synthase
17405240166440316665022100gp: AB023377_1Corynebacterium glutamicum100.0100.0675transketolase
ATCC 31833 tkt
17415241166667316677521080sp: TAL_MYCLEMycobacterium leprae62.085.2358transaldolase
MLCL536.39 tal
17425242166776416666011164
17435243166795016694011452gsp: W27612Brevibacterium flavum99.8100.0484glucose-6-phosphate
dehydrogenase
1744524416694191670375957pir: A70917Mycobacterium tuberculosis40.671.7318oxppcycle protein (glucose 6-
H37Rv Rv1446c opcAphosphate dehydrogenase
assembly protein)
1745524516703951671099705sp: SOL3_YEASTSaccharomyces cerevisiae28.758.12586-phosphogluconolactonase
S288C YHR163W sol3
1746524616716771671273405sp: SAOX_BACSNBacillus sp. NS-12935.257.8128sarcosine oxidase
17475247167172316731231401gp: AF126281_1Rhodococcus erythropolis 24.646.6500transposase (IS1676)
1748524816741051673266840gp: CGL007732_5Corynebacterium glutamicum100.0100.0205sarcosine oxidase
ATCC 13032 soxA
1749524916772111677384174
1750525016787561678070687
1751525116791481680128981
1752525216811081680332777sp: TPIS_CORGLCorynebacterium glutamicum99.299.6259triose-phosphate isomerase
AS019 ATCC 13059 tpiA
1753525316812631681670408SP: YCQ3_YEASTSaccharomyces cerevisiae37.051.0128probable membrane protein
YCR013c
17545254168240416811901215sp: PGK_CORGLCorynebacterium glutamicum98.098.5405phosphoglycerate kinase
AS019 ATCC 13059 pgk
17555255168362516826241002sp: G3P_CORGLCorynebacterium glutamicum99.199.7333glyceraldehyde-3-phosphate
AS019 ATCC 13059 gapdehydrogenase
1756525616850971684117981pir: D70903Mycobacterium tuberculosis63.987.4324hypothetical protein
H37Rv Rv1423
17575257168613216851101023sp: YR40_MYCTUMycobacterium tuberculosis56.382.5309hypothetical protein
H37Rv Rv1422
1758525816870781686152927sp: YR39_MYCTUMycobacterium tuberculosis52.076.2281hypothetical protein
H37Rv Rv1421
17595259168919016871032088sp: UVRC_PSEFLSynechocystis sp. PCC680334.461.5701excinuclease ABC subunit C
uvrC
1760526016897791689201579sp: YR35_MYCTUMycobacterium tuberculosis32.768.7150hypothetical protein
H37Rv Rv1417
1761526116903451689869477SP: RISB_ECOLIEscherichia coli K1243.572.11546,7-dimethyl-8-ribityllumazine
synthase
1762526216906941690921228GSP: Y83273Bacillus subtilis59.068.072polypeptide encoded by rib operon
1763526316907081691421714GSP: Y83272Bacillus subtilis26.048.0217riboflavin biosynthetic protein
1764526416910121691347336GSP: Y83273Bacillus subtilis44.052.0106polypeptide encoded by rib operon
17655265169162516903601266gp: AF001929_1Mycobacterium tuberculosis ribA65.684.7404GTP cyclohydrolase II and 3,4-
dihydroxy-2-butanone 4-phosphate
synthase (riboflavin synthesis)
1766526616922711691639633sp: RISA_ACTPLActinobacillus47.479.2211riboflavin synthase alpha chain
pleuropneumoniae ISU-178 ribE
1767526716932581692275984sp: RIBD_ECOLIEscherichia coli K12 ribD37.362.7365riboflavin-specific deaminase
1768526816939181693262657sp: RPE_YEASTSaccharomyces cerevisiae43.673.1234ribulose-phosphate 3-epimerase
S288C YJL121C rpe1
17695269169529816939671332sp: SUN_ECOLIEscherichia coli K12 sun30.860.7448nucleolar protein NOL1/NOP2
(eukaryotes) family
1770527016964431695499945sp: FMT_PSEAEPseudomonas aeruginosa fmt41.667.9308methionyl-tRNA formyltransferase
1771527116969721696466507sp: DEF_BACSUBacillus subtilis 168 def44.772.7150polypeptide deformylase
17725272169914716970842064sp: PRIA_ECOLIEscherichia coli priA22.946.3725primosomal protein n
17735273170039716991771221gsp: R80060Brevibacterium flavum MJ-23399.399.5407S-adenosylmethionine synthetase
17745274170176717005081260sp: DFP_MYCTUMycobacterium tuberculosis58.080.9409DNA/pantothenate metabolism
H37Rv RV1391 dfpflavoprotein
1775527517023221702032291sp: YD90_MYCTUMycobacterium tuberculosis70.487.781hypothetical protein
H37Rv Rv1390
1776527617030371702411627pir: KIBYGUSaccharomyces cerevisiae guk139.874.7186guanylate kinase
1777527717033081702991318pir: B70899Mycobacterium tuberculosis80.690.3103integration host factor
H37Rv Rv1388 mIHF
1778527817043501703517834sp: DCOP_MYCTUMycobacterium tuberculosis51.873.6276orotidine-5′-phosphate
H37Rv uraAdecarboxylase
17795279170769717043593339pir: SYECCPEscherichia coli carB53.177.51122carbamoyl-phosphate synthase
large chain
17805280170888417077061179sp: CARA_PSEAEPseudomonas aeruginosa45.470.1381carbamoyl-phosphate synthase
ATCC 15692 carAsmall chain
17815281171035717090171341sp: PYRC_BACCLBacillus caldolyticus DSM 40542.867.7402dihydroorotase
pyrC
1782528217113481710413936sp: PYRB_PSEAEPseudomonas aeruginosa48.679.7311aspartate carbamoyltransferase
ATCC 15692
1783528317119271711352576sp: PYRR_BACCLBacillus caldolyticus DSM 40554.080.1176phosphoribosyl transferase or
pyrRpyrimidine operon regulatory protein
17845284171259617137591164sp: Y00R_MYCTUMycobacterium tuberculosis39.773.4297cell division inhibitor
H37Rv Rv2216
1785528517138301714306477
1786528617142991714760462
1787528717147411714950210
1788528817160621715382681sp: NUSB_BACSUBacillus subtilis nusB33.669.3137N utilization substance protein B
(regulation of rRNA biosynthesis by
transcriptional antitermination)
1789528917166921716132561sp: EFP_BRELABrevibacterium lactofermentum97.998.4187elongation factor P
ATCC 13869 efp
17905290171786817167801089gp: AF124600_4Corynebacterium glutamicum99.5100.0217cytoplasmic peptidase
AS019 pepQ
17915291171903217179381095gp: AF124600_3Corynebacterium glutamicum98.699.73613-dehydroquinate synthase
AS019 aroB
1792529217195981719107492gp: AF124600_2Corynebacterium glutamicum100.0100.0166shikimate kinase
AS019 aroK
1793529317213811720971411sp: LEP3_AERHYAeromonas hydrophila tapD35.254.9142type IV prepilin-like protein specific
leader peptidase
1794529417217251721423303gp: SC1A2_22Streptomyces coelicolor A3(2)45.868.783bacterial regulatory protein, arsR
SC1A2.22family
17955295172178017228531074gp: AF109162_2Corynebacterium diphtheriae35.973.2340ABC transporter
hmuU
1796529617228071722202606
1797529717228701723826957pir: A75169Pyrococcus abyssi Orsay23.650.7373iron(III) ABC transporter,
PAB0349periplasmic-binding protein
1798529817238261724578753sp: FHUC_BACSUBacillus subtilis 168 fhuC38.371.7230ferrichrome transport ATP-binding
protein
1799529917254391724612828pir: D70660Mycobacterium tuberculosis50.060.0259shikimate 5-dehydrogenase
H37Rv aroE
18005300172662517254591167pir: E70660Mycobacterium tuberculosis41.870.1395hypothetical protein
H37Rv Rv2553c
1801530117271701726625546pir: F70660Mycobacterium tuberculosis52.869.6161hypothetical protein
H37Rv Rv2554c
18025302173004817273852664sp: SYA_THIFEThiobacillus ferrooxidans ATCC43.371.8894alanyl-tRNA synthetase
33020 alaS
18035303173154217301661377sp: Y0A9_MYCTUMycobacterium tuberculosis65.484.8454hypothetical protein
H37Rv Rv2559c
18045304173282217315991224
18055305173481117329881824sp: SYD_MYCLEMycobacterium leprae aspS71.189.2591aspartyl-tRNA synthetase
1806530617350561735946891sp: Y0BQ_MYCTUMycobacterium tuberculosis46.174.1297hypothetical protein
H37Rv Rv2575
18075307173867917360042676sp: AMYH_YEASTSaccharomyces cerevisiae26.153.6839glucan 1,4-alpha-glucosidase
S288C YIR019C sta1
18085308174056917387131857sp: YHGE_BACSUBacillus subtilis yhgE23.154.0742phage infection protein
1809530917412191740572648
1810531017413131741906594gp: SCE68_13Streptomyces coelicolor A3(2)29.262.0192transcriptional regulator
SCE68.13
1811531117418931742606714
18125312174270117438131113gp: SCE15_13Streptomyces coelicolor A3(2)72.888.1371oxidoreductase
SCE15.13c
1813531317438431743968126
1814531417440251744519495sp: SLFA_PSEAEPseudomonas aeruginosa PAO137.177.6116NADH-dependent FMN reductase
slfA
18155315174488417462301347sp: SDHL_ECOLIEscherichia coli K12 sdaA46.871.4462L-serine dehydratase
1816531617467281747588861
18175317174791817462331686prf: 2423362AEnterococcus casseliflavus glpO28.453.9598alpha-glycerolphosphate oxidase
18185318174927617479901287sp: SYH_STAAUStaphylococcus aureus 43.272.2421histidyl-tRNA synthetase
SR17238 hisS
1819531917499631749325639gp: CJ11168X3_127Campylobacter jejuni 40.362.1211hydrolase
NCTC11168 Cj0809c
1820532017504271750933507prf: 2313309AStreptomyces chrysomallus35.461.1175cyclophilin
sccypB
1821532117509641751200237
1822532217514971752051555gp: AF038651_4Corynebacterium glutamicum98.4100.0128hypothetical protein
ATCC 13032 orf4
1823532317521861752527342
18245324175489417526152280gp: AF038651_3Corynebacterium glutamicum99.999.9760GTP pyrophosphokinase
ATCC 13032 rel
1825532517554791754925555gp: AF038651_2Corynebacterium glutamicum99.5100.0185adenine phosphoribosyltransferase
ATCC 13032 apt
1826532617557481755599150gp: AF038651_1Corynebacterium glutamicum98.098.849dipeptide transport system
ATCC 13032 dciAE
18275327175722817554861743sp: Y0BG_MYCTUMycobacterium tuberculosis30.760.9558hypothetical protein
H37Rv Rv2585c
18285328175879717575891209sp: SECF_ECOLIEscherichia coli K12 secF25.957.2332protein-export membrane protein
1829532917597071760336630
18305330176073417588031932prf: 2313285ARhodobacter capsulatus secD24.452.0616protein-export membrane protein
1831533117613671761005363sp: Y0BD_MYCLEMycobacterium leprae39.666.0106hypothetical protein
MLCB1259.04
18325332176249817614191080sp: RUVB_ECOLIEscherichia coli K12 ruvB55.381.9331holliday junction DNA helicase
1833533317631341762517618sp: RUVA_MYCLEMycobacterium leprae ruvA45.274.3210holliday junction DNA helicase
1834533417638391763177663sp: RUVC_ECOLIEscherichia coli K12 ruvC35.663.3180crossover junction
endodeoxyribonuclease
1835533517647421763990753sp: YEBC_ECOLIEscherichia coli K12 ORF24649.278.4250hypothetical protein
yebC
1836533617658601765015846sp: TESB_ECOLIEscherichia coli K12 tesB38.568.6283acyl-CoA thiolesterase
1837533717659691766442474gp: SC10A5_9Streptomyces coelicolor A3(2)31.561.3111hypothetical protein
SC10A5.09c
1838533817669481766487462pir: H70570Mycobacterium tuberculosis38.261.2170hypothetical protein
H37Rv Rv2609c
18395339176803017669481083sp: GPI3_YEASTSaccharomyces cerevisiae21.749.3414hexosyltransferase or N-
S288C spt14acetylglucosaminyl-
phosphatidylinositol biosynthetic
protein
1840534017689961768034963gp: SCL2_16Streptomyces coelicolor A3(2)46.467.8295acyltransferase
SCL2.16c
1841534117696781769022657pir: C70571Mycobacterium tuberculosis48.278.078CDP-dlacylglycerol—glycerol-3-
H37Rv Rv2612c pgsAphosphate phosphatidyltransferase
1842534217703401769681660pir: D70571Mycobacterium tuberculosis54.678.4194histidine triad (HIT) family protein
H37Rv Rv2613c
18435343177238417703272058sp: SYT2_BACSUBacillus subtilis thrZ42.068.9647threonyl-tRNA synthetase
18445344177386317726581206sp: YWBN_BACSUBacillus subtilis ywbN34.361.8400hypothetical protein
1845534517738811774444564
1846534617744381773893546
1847534717751911774457735
1848534817772691777646378
1849534917774441778037594
18505350177950817781021407
1851535117801681779554615
1852535217809051780507399
1853535317815851781019567sp: PUAC_STRLPStreptomyces anulatus pac36.364.2190puromycin N-acetyltransferase
18545354178170517827901086
18555355178328117843811101
1856535617840801783382699
18575357178547317828942580
18585358178684417857321113
18595359178882917869071923
1860536017890801789562483
1861536117895801789768189
1862536217897461790057312
1863536317908891790461429
1864536417918421792438597sp: AFUC_ACTPLActinobacillus28.728.7202ferric transport ATP-binding protein
pleuropneumoniae afuC
1865536517924281793426999
1866536617936541793496159
18675367179371417948201107
1868536817952021795621420
1869536917955911796181591gp: AF088896_20Zymomonas mobilis dfp27.166.7129pantothenate metabolism
flavoprotein
1870537017961861797049864
1871537117973501797769420
1872537217979691797850120
1873537317987571798023735
1874537417991821799406225
1875537517994731800366894
1876537618006041800449156
1877537718008341801307474
1878537818013441802096753
1879537918025771802155423
1880538018027331803419687
1881538118034651803893429
1882538218041341804598465
1883538318046291804865237
1884538418049191805599681
1885538518057271806686960
1886538618069171807396480
1887538718074331808113681
1888538818081371808421285
1889538918084581808832375
1890539018097611810372612sp: TNP2_ECOLIEscherichia coli tnpR51.178.0186transposon TN21 resolvase
18915391181054118115451005
1892539218115641811938375
1893539318122151812691477sp: PVH1_YEASTSaccharomyces cerevisiae29.351.8164protein-tyrosine phosphatase
S288C YIR026C yvh1
1894539418128811813606726
1895539518128821812460423
1896539618137801814517738gp: SCA32WHIH_6Streptomyces coelicolor A3(2)34.365.7216sporulation transcription factor
whiH
1897539718148631815651789
1898539818156731816128456
1899539918164511816636186
1900540018171321817803672
1901540118178031818219417
1902540218184601818774315
1903540318187981819166369
1904540418199541819748207
19055405182238218201812202pir: C72285Thermotoga maritima MSB822.655.2545hypothetical protein
TM1189
19065406182257718243221746
1907540718243711824589219
1908540818247841824927144
1909540918256061825178429
1910541018260241826557534PIR: S60891Corynebacterium glutamicum63.075.0166hypothetical protein
1911541118266441825751894pir: S60890Corynebacterium glutamicum87.995.6298insertion element (IS3 related)
orf2
1912541218269371826644294pir: S60889Corynebacterium glutamicum72.384.2101insertion element (IS3 related)
orf1
1913541318299001829688213
19145414183076518320631299
19155415183216718340441878sp: RECJ_ERWCHErwinia chrysanthemi recJ24.050.6622single-stranded-DNA-specific
exonuclease
1916541618349281834149780
19175417183667518383241650pir: T13302Streptococcus phage phi-O120531.864.3381primase
ORF13
19185418183834918421373789
1919541918422351842681447
1920542018428041843337534
19215421184351818453561839sp: Y018_MYCPNMycoplasma pneumoniae ATCC22.144.7620helicase
29342 yb95
1922542218454831845857375
1923542318458721846207336pir: T13144Bacteriophage N15 gene5736.764.2109phage N15 protein gp57
1924542418466981846333366
1925542518473151847932618
1926542618479381848474537
1927542718485091849036528
1928542818489881849785798
1929542918497811849966186
1930543018500351850406372
1931543118504151849978438
1932543218510491850474576
19335433185122018524401221gp: SPAPJ760_2Schizosaccharomyces pombe28.749.8422actin binding protein with SH3
SPAPJ760.02cdomains
1934543418514731852324852
19355435185247918538731395
1936543618542611854854594
1937543718550581855237180
19385438185553218567881257gp: SC5C7_14Streptomyces coelicolor23.652.5347ATP/GTP binding protein
SC5C7.14
19395439185688518587381854
19405440185876318607271965sp: CLPA_ECOLIEscherichia coli K12 clpA30.261.0630ATP-dependent Clp proteinase ATP-
binding subunit
1941544118607521861225474
1942544218613201861475156
1943544318618421861519324
1944544418620881862399312
19455445186294518652992355sp: PCRA_STAAUStaphylococcus aureus SA2021.445.9693ATP-dependent helicase
pcrA
1946544618652651865822558
1947544718658421866219378
1948544818663281866792465
1949544918668321867095264
1950545018670981867874777gp: SCH17_7Streptomyces coelicolor A3(2)25.947.8224hypothetical protein
SCH17.07c
1951545118678861868587702prf: 2514444YBacteriophage phi-C31 gp5231.761.5208deoxynucleotide monophosphate
kinase
1952545218688951868671225
19535453187109218689272166
1954545418713731871101273
19555455187788618713806507
19565456187831218794001089prf: 2403350ACorynebacterium glutamicum99.299.7363type II 5-cytosoine
ATCC 13032 cglIMmethyltransferase
19575457187941218804851074pir: A55225Corynebacterium glutamicum99.799.7358type II restriction endonuclease
ATCC 13032 cglIR
19585458188399018824701521
1959545918849361884220717
19605460188523018870471818gp: SC1A2_16Streptomyces coelicolor A3(2)24.645.8504hypothetical protein
SC1A2.16c
1961546118874051887590186
1962546218880381887688351gp: AE001973_4Deinococcus radiodurans46.770.090SNF2/Rad54 helicase-related
DR1258protein
1963546318890941888231864pir: T13226Lactobacillus phage phi-gle33.156.4163hypothetical protein
Rorf232
1964546418895301889859330
19655465189170718900281680gp: AF188935_16Bacillus anthracis pXO2-1620.747.9537hypothetical protein
19665466189303718918321206
19675467189468018933881293
19685468189723118947392493
19695469189915818973741785sp: CLPB_ECOLIEscherichia coli clpB25.352.5724endopeptidase Clp ATP-binding
chain B
1970547018998531899233621
19715471190091618998041113
1972547219019111901066846
1973547319019751902955981
1974547419028831902005879
1975547519030281903225198
19765476190587819031132766pir: S23647Homo sapiens numA20.149.11004nuclear mitotic apparatus protein
1977547719065721905973600
19785478190791419066641251
1979547919086601907965696
1980548019094981908785714
19815481191050819095011008
19825482191230019106421659
19835483191382019123331488
1984548419143711913973399
19855485191623319147251509
1986548619163741916733360
1987548719169441917165222
1988548819176401917329312
1989548919182081917564645
1990549019194611918703759
1991549119201941919646549
1992549219212761920347930
1993549319253901925695306
1994549419256821926038357
19955495192601019215474464pir: T03099Sus scrofa domestica23.249.21408submaxillary apomucin
1996549619268371926259579
1997549719281891927245945
1998549819282111928381171sp: MTE1_ECOLIEscherichia coli ecoR142.665.661modification methylase
1999549919285341928908375
20005500193087919290591821
2001550119311901930990201
2002550219318881931421468
2003550319323151931935381pir: H70638Mycobacterium tuberculosis38.658.8114hypothetical protein
H37Rv Rv1956
2004550419328791932373507
2005550519343581933522837
2006550619359121934971942sp: Y137_METJAMethanococcus jannaschii27.154.6328hypothetical protein
MJ0137
2007550719362261936849624
2008550819372021937411210
2009550919380191937486534
20105510193894519401351191
2011551119390641938531534
2012551219402571940844588
2013551319411071941550444
2014551419424841941732753
2015551519425101942812303
2016551619430951943310216
2017551719433451943653309
2018551819436801944564885
2019551919454351944608828prf: 2509434AEnterococcus faecalis esp23.044.1304surface protein
2020552019458911945595297
2021552119463321945952381
2022552219470371946609429
20235523194865019470701581sp: CSP1_CORGLCorynebacterium glutamicum30.754.4270major secreted protein PS1 protein
(Brevibacterium flavum) ATCCprecursor
17965 csp1
20245524195145019490212430
2025552519524851951619867
20265526195482219525462277sp: TOP3_ECOLIEscherichia coli topB23.850.9597DNA topoisomerase III
20275527195828719562032085
2028552819593401958450891
2029552919601961959765432
2030553019611141960371744
20315531196300019611141887sp: CSP1_CORGLCorynebacterium glutamicum29.754.7344major secreted protein PS1 protein
(Brevibacterium flavum) ATCCprecursor
17965 csp1
2032553219634291963139291
20335533196474319635141230
20345534196590219647271176
2035553519662671965911357
2036553619663011966984684sp: NUC_STAAUStaphylococcus aureus nuc30.457.7227thermonuclease
2037553719674351967289147
2038553819676041968167564
20395539196826419697151452
2040554019697451970203459
20415541197025419714741221
20425542197167219730901419
2043554319731471973737591
2044554419738091974204396
2045554519742671974503237
2046554619751711975794624prf: 2313347BShewanella sp. ssb24.959.1225single stranded DNA-binding protein
2047554719759161976494579
2048554819765221976983462
2049554919770431977549507
2050555019777421978329588
2051555119783891978721333
2052555219786601979217558
2053555319792391979808570
2054555419799741980885912sp: S24D_ANOGAAnopheles gambiae AgSP24D25.752.6249serine protease
2055555519809651981657693
2056555619816631982028366
2057555719820711982817747
2058555819820911981912180
2059555919831861983548363
2060556019836111983883273
2061556119839181984181264
2062556219842171984450234
2063556319843871984728342
2064556419850921985364273
2065556519853731985071303
20665566198659019854421149sp: VINT_BPML5Mycobacterium phage L5 int29.655.9406integrase
2067556719878961987507390gsp: R23011Brevibacterium lactofermentum83.994.4124transposase (divided)
CGL2005 ISaB1
2068556819883031987887417gsp: R23011Brevibacterium lactofermentum70.984.6117transposase (divided)
CGL2005 ISaB1
2069556919883831988589207
2070557019884831988370114gsp: R21601Brevibacterium lactofermentum80.796.831transposition repressor
CGL2005 ISaB1
2071557119886641988530135pir: S60889Corynebacterium glutamicum74.488.443insertion element (IS3 related)
orf1
2072557219896051988778828gp: SCJ11_12Streptomyces coelicolor A3(2)31.153.7270transposase
SCJ11.12
2073557319906671991020354
2074557419907641989874891
2075557519916201991189432
2076557619925381991795744
20775577199412119925381584sp: CSP1_CORGLCorynebacterium glutamicum25.037.0153major secreted protein PS1 protein
(Brevibacterium flavum) ATCCprecursor
17965 csp1
2078557819952941994608687sp: VINT_BPML5Mycobacterium phage L5 int28.756.1223integrase
2079557919960881995783306pir: F64546Helicobacter pylori 2669539.876.188sodium-dependent transporter
HP0214
2080558019961061996537432sp: YXAA_BACSUBacillus subtilis yxaA48.981.592hypothetical protein
2081558119967681997112345
2082558219971681997503336
2083558319975451998240696pir: C70968Mycobacterium tuberculosis33.564.4233riboflavin biosynthesis protein
H37Rv Rv2671 ribD
20845584199828919995421254pir: E70968Mycobacterium tuberculosis42.571.9384potential membrane protein
H37Rv Rv2673
2085558519995421999949408gp: AF128264_2Streptococcus gordonii msrA41.367.5126methionine sulfoxide reductase
2086558620001321999707426
2087558720012162000521696pir: H70968Mycobacterium tuberculosis55.277.2232hypothetical protein
H37Rv Rv2676c
2088558820014892002112624pir: C70528Mycobacterium tuberculosis55.778.6201hypothetical protein
H37Rv Rv2680
20895589200207220033341263sp: RND_HAEINHaemophilus influenzae Rd25.952.8371ribonuclease D
KW20 HI0390 rnd
20905590200530920034021908gp: AB026631_1Streptomyces sp. CL190 dxs55.378.56181-deoxy-D-xylulose-5-phosphate
synthase
20915591200669720054621236pir: E72298Thermotoga maritima MSB825.452.3472RNA methyltransferase
TM1094
2092559220066982006979282
2093559320076372006777861pir: C70530Mycobacterium tuberculosis38.162.7268hypothetical protein
H37Rv Rv2696c
2094559420081842007738447sp: DUT_STRCOStreptomyces coelicolor A3(2)55.082.1140deoxyuridine 5′-triphosphate
SC2E9.09 dutnucleotidohydrolase
2095559520082502008798549pir: E70530Mycobacterium tuberculosis46.070.7150hypothetical protein
H37Rv Rv2698
2096559620090822008876207
2097559720095702009280291pir: F70530Mycobacterium tuberculosis58.081.0100hypothetical protein
H37Rv Rv2699c
2098559820105392009724816sp: SUHB_ECOLIEscherichia coli K12 suhB38.468.2198extragenic suppressor protein
2099559920105552011382828sp: PPGK_MYCTUMycobacterium tuberculosis54.480.2248polyphosphate glucokinase
H37Rv RV2702 ppgK
21005600201186320133561494prf: 2204286ACorynebacterium glutamicum98.098.6500sigma factor or RNA polymerase
sigAtranscription factor
21015601201549620141621335sp: YRKO_BACSUBacillus subtilis yrkO23.951.4422hypothetical membrane protein
2102560220161212015585537
21035603201796620162571710sp: Y065_MYCTUMycobacterium tuberculosis61.380.8578hypothetical protein
H37Rv Rv2917
2104560420181192018754636pir: H70531Mycobacterium tuberculosis32.359.1127hypothetical membrane protein
H37Rv Rv2709
2105560520182022017966237pir: G70531Mycobacterium tuberculosis65.885.576hypothetical protein
H37Rv Rv2708c
21065606201874420202761533gp: SCH5_8Streptomyces coelicolor A3(2)33.561.2523transferase
SCH5.08c
2107560720202932020724432prf: 2204286CCorynebacterium glutamicum97.2100.0144hypothetical protein
ATCC 13869 ORF1
2108560820222662022949684pir: I40339Corynebacterium glutamicum98.799.6228iron dependent repressor or
ATCC 13869 dtxRdiphtheria toxin repressor
2109560920225462022313234GP: AF010134_1Streptomyces aureofaciens62.064.077putative sporulation protein
2110561020229592023945987sp: GALE_BRELACorynebacterium glutamicum99.199.1329UDP-glucose 4-epimerase
ATCC 13869 (Brevibacterium
lactofermentum) galE
21115611202527020239481323
2112561220254232026379957pir: E70532Mycobacterium tuberculosis45.379.0305hypothetical protein
H37Rv Rv2714
21135613202649420290432550sp: MTR4_YEASTSaccharomyces cerevisiae24.450.7661ATP-dependent RNA helicase
YJL050W dob1
2114561420291772030157981sp: OXYR_ECOLIEscherichia coli oxyR35.865.6299hydrogen peroxide-inducible genes
activator
21155615203136520302771089
21165616203147820353833906sp: HRPA_ECOLIEscherichia coli hrpA49.276.21298ATP-dependent helicase
2117561720358802035431450gp: SCAJ4870_3Streptomyces clavuligerus nrdR61.486.2145regulatory protein
2118561820364092035990420
2119561920368122037507696sp: LEXA_BACSUBacillus subtilis dinR46.971.6222SOS regulatory protein
2120562020378152038591777sp: GATR_ECOLIEscherichia coli K12 gatR33.967.8245galactitol utilization operon repressor
2121562120385912039550960gp: SCE22_14Streptomyces coelicolor A3(2)27.255.6320phosphofructokinase (fructose 1-
SCE22.14cphosphate kinase)
21225622204132120396181704sp: PT1_BACSTBacillus stearothermophilus ptsI34.364.0592phosphoenolpyruvate-protein
phosphotransferase
2123562320417282042519792sp: GLPR_ECOLIEscherichia coli K12 glpR26.762.6262glycerol-3-phosphate regulon
repressor
2124562420425192043508990sp: K1PF_RHOCARhodobacter capsulatus fruK33.055.73451-phosphofructokinase or 6-
phosphofructokinase
21255625204373620455711836sp: PTFB_ECOLIEscherichia coli K12 fruA43.069.6549PTS system, fructose-specific IIBC
component
2126562620457622046028267sp: PTHP_BACSTBacillus stearothermophilus XL-37.071.681phosphocarrier protein
65-6 ptsH
2127562720472952046714582
21285628204860620473201287sp: PYRP_BACCLBacillus caldolyticus pyrP39.170.5407uracil permease
21295629205010720486501458gp: AF145049_8Streptomyces fradiae orf11*54.480.0419ATP/GTP-binding protein
2130563020503212051106786
2131563120513062051842537
2132563220526752051845831sp: DAPF_HAEINHaemophilus influenzae Rd33.564.7269diaminopimelate epimerase
KW20 HI0750 dapF
2133563320535862052684903sp: MIAA_ECOLIEscherichia coli K12 miaA40.068.7300tRNA delta-2-
isopentenylpyrophosphate
transferase
2134563420542832053609675
21355635205440320557611359pir: B70506Mycobacterium tuberculosis48.575.7445hypothetical protein
H37Rv Rv2731
21365636205574320547241020
21375637205576520567871023
2138563820577882057120669pir: C70506Mycobacterium tuberculosis29.063.7190hypothetical membrane protein
H37Rv Rv2732c
21395639205942020578551566sp: Y195_MYCLEMycobacterium leprae68.486.4494hypothetical protein
B2235_C2_195
2140564020597742060499726sp: GLUA_CORGLCorynebacterium glutamicum99.699.6242glutamate transport ATP-binding
ATCC 13032 gluAprotein
2141564120604142060196219GSP: Y75358Neisseria gonorrhoeae66.073.071Neisserial polypeptides predicted to
be useful antigens for vaccines and
diagnostics
2142564220616292062312684sp: GLUC_CORGLCorynebacterium glutamicum100.0100.0225glutamate transport system
ATCC 13032 gluCpermease protein
2143564320624412063259819sp: GLUD_CORGLCorynebacterium glutamicum99.399.6273glutamate transport system
(Brevibacterium flavum) ATCCpermease protein
13032 gluD
2144564420638942063298597sp: RECX_MYCLEMycobacterium leprae recX34.566.9142regulatory protein
2145564520656272065394234pir: A70878Mycobacterium tuberculosis40.371.667hypothetical protein
H37Rv Rv2738c
2146564620664042065667738
2147564720665662067141576sp: BIOY_BACSHBacillus sphaericus bioY33.061.4197biotin synthase
2148564820671682067866699sp: POTG_ECOLIEscherichia coli K12 potG33.269.5223putrescine transport ATP-binding
protein
2149564920678662068474609pir: F69742Bacillus subtilis ybaF24.658.8228hypothetical membrane protein
2150565020687032069392690pir: B60176Mycobacterium tuberculosis41.778.5228hypothetical protein
2151565120693832068556828sp: 35KD_MYCTUMycobacterium tuberculosis72.589.6269hypothetical protein (35 kD protein)
H37Rv RV2744C
2152565220699362069616321pir: H70878Mycobacterium tuberculosis54.278.383regulator (DNA-binding protein)
H37Rv Rv2745c
2153565320705122069997516sp: CINA_STRPNStreptococcus pneumoniae R6X41.868.5165competence damage induced
cinAproteins
2154565420711212070519603prf: 2421334DStreptococcus pyogenes pgsA38.872.5160phosphotidylglycerophosphate
synthase
2155565520713152071599285pir: T10688Arabidopsis thaliana24.852.1117hypothetical protein
ATSP: T16|18.20
2156565620716242071740117gp: AF071810_1Streptococcus pneumoniae60.070.030surface protein (Peumococcal
DBL5 pspAsurface protein A)
2157565720720662072878813
21585658207290520717991107prf: 2119295DEscherichia coli terC31.059.8358tellurite resistance protein
21595659207605620732942763sp: SP3E_BACSUBacillus subtilis 168 spoIIIE38.064.6845stage III sporulation protein E
2160566020770242076392633gp: SC4G6_14Streptomyces coelicolor A3(2)33.361.0216hypothetical protein
SC4G6.14
21615661207927520771222154sp: YOR4_CORGLCorynebacterium glutamicum99.199.4645hypothetical protein
ATCC 13032 orf4
2162566220811362080387750sp: YDAP_BRELACorynebacterium glutamicum99.299.6250hypothetical protein
(Brevibacterium lactofermentum)
ATCC 13869 orf2
2163566320821152082813699
2164566420823682082105264
21655665208519020829322259prf: 2217311AStreptomyces antibioticus gpsI65.485.3742guanosine pentaphosphate
synthetase
2166566620857022085436267pir: F69700Bacillus subtilis rpsO64.088.88930S ribosomal protein S15
2167566720868262085879948prf: 2518365ALeishmania major35.163.3319nucleoside hydrolase
21685668208794120869191023sp: RIBF_CORAMCorynebacterium56.279.0329bifunctional protein (riboflavin kinase
ammoniagenes ATCC 6872 ribFand FAD synthetase)
2169566920879732088863891sp: TRUB_BACSUBacillus subtilis 168 truB32.761.7303tRNA pseudouridine synthase B
2170567020881812087954228PIR: PC4007Corynebacterium65.073.047hypothetical protein
ammoniagenes
2171567120898682089218651gp: SC5A7_23Streptomyces coelicolor A3(2)42.262.5237hypothetical protein
SC5A7.23
2172567220906642089861804pir: B70885Mycobacterium tuberculosis46.968.9273phosphoesterase
H37Rv Rv2795c
21735673209205520907511305pir: G70693Mycobacterium tuberculosis51.078.8433DNA damaged inducible protein f
H37Rv Rv2836c dinF
2174567420930462092051996pir: H70693Mycobacterium tuberculosis36.770.8308hypothetical protein
H37Rv Rv2837c
2175567520935012093055447sp: RBFA_BACSUBacillus subtilis 168 rbfA32.470.4108ribosome-binding factor A
21765676209672320937123012sp: IF2_STIAUStigmatella aurantiaca DW4 infB37.762.91103translation initiation factor IF-2
2177567720971792096844336gp: SC5H4_29Streptomyces coelicolor A3(2)44.666.383hypothetical protein
SC5H4.29
2178567820983752097380996sp: NUSA_BACSUBacillus subtilis 168 nusA42.371.0352n-utilization substance protein
(transcriptional
termination/antitermination factor)
21795679209856220998151254
2180568020989452098412534pir: E70588Mycobacterium tuberculosis34.665.5165hypothetical protein
H37Rv Rv2842c
21815681210024021018411602sp: DPPE_BACSUBacillus subtilis 168 dppE25.360.9534peptide-binding protein
2182568221020232102946924sp: DPPB_ECOLIEscherichia coli K12 dppB37.769.4337peptidetransport system permease
2183568321029752103973999prf: 1709239CBacillus subtilis spo0KC38.469.2292oligopeptide permease
21845684210397321057031731pir: H70788Mycobacterium tuberculosis57.681.3552peptidetransport system ABC-
H37Rv Rv3663c dppDtransporter ATP-binding protein
21855685210756421058011764sp: SYP_MYCTUMycobacterium tuberculosis67.084.6578prolyl-tRNA synthetase
H37Rv Rv2845c proS
2186568621076522108386735gp: SCC30_5Streptomyces coelicolor A3(2)39.565.0243hypothetical protein
SCC30.05
2187568721091472108389759sp: BCHD_RHOSHRhodobacter sphaeroides ATCC32.460.737magnesium-chelatase subunit
17023 bchD
21885688211025521091551101prf: 2503462AAHeliobacillus mobilis bchI46.569.6342magnesium-chelatase subunit
2189568921111832110434750prf: 2108318BPropionibacterium freudenreichii49.073.8237uroporphyrinogen III
cobAmethyltransferase
21905690211123821126591422sp: YPLC_CLOPEClostridium perfringens NCIB41.268.7488hypothetical protein
10662 ORF2
2191569121136162112717900gp: SC5H1_10Streptomyces coelicolor A3(2)35.162.3151hypothetical protein
SC5H1.10c
21925692211576121167741014pir: A70590Mycobacterium tuberculosis37.665.7338hypothetical protein
H37Rv Rv2854
21935693211691621183101395sp: GSHR_BURCEBurkholderia cepacia AC110053.076.6466glutathione reductase
gor
2194569421179562117015942
2195569521186072119080474
2196569621191392119495357
2197569721196282120356729
2198569821211472120359789sp: AMPM_ECOLIEscherichia coli K12 map47.275.8252methionine aminopeptidase
21995699212316121212961866prf: 2224268AStreptomyces clavuligerus pcbR27.356.5630penicillin binding protein
2200570021238482123219630prf: 2518330BCorynebacterium diphtheriae44.072.2216response regulator (two-component
chrAsystem response regulator)
22015701212499621238481149prf: 2518330ACorynebacterium diphtheriae29.556.8424two-component system sensor
chrShistidine kinase
2202570221250892126045957gp: AE001863_70Deinococcus radiodurans24.458.1360hypothetical membrane protein
DRA0279
2203570321260642126753690prf: 2420410PBacillus subtilis 168 yvrO37.371.1225ABC transporter
2204570421270872126926162
22055705212848321273501134sp: GCPE_ECOLIEscherichia coli K12 gcpE44.373.8359hypothetical protein (gcpE protein)
2206570621288502129461612
22075707212988021286691212pir: G70886Mycobacterium tuberculosis43.073.6405hypothetical membrane protein
H37Rv Rv2869c
2208570821303062130950645GSP: Y37145Chlamydia trachomatis36.043.0147polypeptides can be used as
vaccines against Chlamydia
trachomatis
22095709213107821299031176sp: DXR_ECOLIEscherichia coli K12 dxr22.842.03121-deoxy-D-xylulose-5-phosphate
reductoisomerase
2210571021313222131762441
2211571121317262131247480
22125712213340221318251578
2213571321342602133406855pir: B72334Thermotoga maritima MSB837.175.1245ABC transporter ATP-binding protein
TM0793
22145714213555121344541098sp: YS80_MYCTUMycobacterium tuberculosis66.078.0356pyruvate formate-lyase 1 activating
H37Rvenzyme
2215571521358842136141258pir: A70801Mycobacterium tuberculosis41.574.594hypothetical membrane protein
H37Rv Rv3760
2216571621370892136235855sp: CDSA_PSEAEPseudomonas aeruginosa33.356.5294phosphatidate cytidylyltransferase
ATCC 15692 cdsA
2217571721378402137286555sp: RRF_BACSUBacillus subtilis 168 frr47.084.3185ribosome recycling factor
2218571821386642137936729prf: 2510355CPseudomonas aeruginosa pyrH28.443.1109uridylate kinase
2219571921389942139854861
2220572021398272139003825sp: EFTS_STRCOStreptomyces coelicolor A3(2)49.676.8280elongation factor Ts
SC2E1.42 tsf
2221572121408862140071816pir: A69699Bacillus subtilis rpsB54.783.525430S ribosomal protein S2
2222572221412572141760504sp: YS91_MYCTUMycobacterium tuberculosis46.058.0120hypothetical protein
H37Rv Rv2891
2223572321426862141763924prf: 2417318AProteus mirabilis xerD40.168.7297site-specific recombinase
22245724214406621428851182sp: YX27_MYCTUMycobacterium tuberculosis39.866.8395hypothetical protein
H37Rv Rv2896c
22255725214558621440661521sp: YX28_MYCTUMycobacterium tuberculosis46.675.8504Mg(2+) chelatase family protein
H37Rv Rv2897c
2226572621459412145576366sp: YX29_MYCTUMycobacterium tuberculosis40.372.3119hypothetical protein
H37Rv Rv2898c
2227572721465662146264303sp: YT01_MYCTUMycobacterium tuberculosis68.396.0101hypothetical protein
H37Rv Rv2901c
2228572821471922146566627sp: RNH2_HAEINHaemophilus influenzae Rd42.669.5190ribonuclease HII
HI1059 rnhB
2229572921472312148022792
2230573021480462147261786prf: 2514288HStreptomyces lividans TK2132.361.1285signal peptidase
sipY
2231573121482312149166936prf: 2510361AStaphylococcus aureus sirA25.459.1323Fe-regulated protein
2232573221495712149359213
2233573321499722149634339sp: RL19_BACSTBacillus stearothermophilus rplS70.388.311150S ribosomal protein L19
2234573421503352150997663sp: THIE_BACSUBacillus subtilis 168 thiE28.460.9225thiamine phosphate
pyrophosphorylase
22355735215103921521181080gp: SC6E10_1Streptomyces coelicolor A3(2)34.064.1376oxidoreductase
SC6E10.01
2236573621521352152329195sp: THIS_ECOLIEscherichia coli K12 thiS37.174.262thiamine biosynthetic enzyme thiS
(thiG1) protein
2237573721523342153113780sp: THIG_ECOLIEscherichia coli K12 thiG48.276.9251thiamine biosynthetic enzyme thiG
protein
22385738215305821541911134prf: 2417383AEmericella nidulans cnxF30.256.8437molybdopterin biosynthesis protein
22395739215673321544602274sp: TEX_BORPEBordetella pertussis TOHAMA I56.678.7776transcriptional accessory protein
tex
2240574021577212156747975pir: A36940Bacillus subtilis 168 degA27.065.3334sporulation-specific degradation
regulator protein
22415741215918121577541428pir: H72105Chlamydophila pneumoniae45.878.3456dicarboxylase translocator
CWL029 ybhl
2242574221592372159019219prf: 2108268ASpinacia oleracea chloroplast40.080.0652-oxoglutarate/malate translocator
22435743216053721592871251sp: PCAB_PSEPUPseudomonas putida pcaB39.166.33503-carboxy-cis, cis-muconate
cycloisomerase
224457442160670216076899
2245574521615032161111393
2246574621621962161507690
2247574721630142162196819sp: TRMD_ECOLIEscherichia coli K12 trmD34.864.8273tRNA (guanine-N1)-
methyltransferase
2248574821630982163745648gp: SCF81_27Streptomyces coelicolor A3(2)30.557.6210hypothetical protein
SCF81.27
2249574921642602163748513sp: RIMM_MYCLEMycobacterium leprae52.372.117216S rRNA processing protein
MLCB250.34. rimM
2250575021643902164737348pir: B71881Helicobacter pylori J99 jhp083929.066.769hypothetical protein
2251575121653092164815495pir: C47154Bacillus subtilis 168 rpsP47.079.58330S ribosomal protein S16
2252575221655232166098576pir: T14151Mus musculus inv32.161.7196inversin
2253575321669902166124867prf: 2512328GStreptococcus agalactiae cylB26.669.1256ABC transporter
2254575421678652166990876prf: 2220349CPyrococcus horikoshll OT3 mtrA35.563.8318ABC transporter
22555755216958421679441641sp: SR54_BACSUBacillus subtilis 168 ffh58.778.2559signal recognition particle protein
2256575621704262171058633
2257575721717152172131417
2258575821722092172877669
22595759217528821737591530sp: FTSY_ECOLIEscherichia coli K12 ftsY37.066.1505cell division protein
2260576021760462175888159
2261576121764022177103702
22625762217950221761103393sp: AMYH_YEASTSaccharomyces cerevisiae22.446.21144glucan 1,4-alpha-glucosidase or
S288C YIR019C sta1glucoamylase S1/S2 precursor
2263576321809182181880963
22645764218309221796283465sp: Y06B_MYCTUMycobacterium tuberculosis48.372.61206chromosome segregation protein
H37Rv Rv2922c smc
2265576521833912183110282sp: ACYP_MYCTUMycobacterium tuberculosis51.173.992acylphosphatase
H37Rv RV2922.1C
22665766218525821834051854
2267576721862082185351858sp: YFER_ECOLIEscherichia coli K12 yfeR23.960.0305transcriptional regulator
2268576821862992187129831pir: S72748Mycobacterium leprae39.373.5257hypothetical membrane protein
MLCL581.28c
2269576921871602187342183
2270577021876792187233447
2271577121883062187692615gp: DNINTREG_3Dichelobacter nodosus gep46.876.6188cation efflux system protein
2272577221891702188313858sp: FPG_ECOLIEscherichia coli K12 mutM or36.166.7285formamidopyrimidine-DNA
fpgglycosylase
2273577321899062189166741pir: B69693Bacillus subtilis 168 rncS40.376.5221ribonuclease III
2274577421904392189906534sp: Y06F_MYCTUMycobacterium tuberculosis35.862.5176hypothetical protein
H37Rv Rv2926c
2275577521913282190540789sp: Y06G_MYCTUMycobacterium tuberculosis50.076.9238hypothetical protein
H37Rv Rv2927c
22765776219152221931651644prf: 2104260GStreptomyces verticillus28.355.6559transport protein
22775777219316521946941530sp: CYDC_ECOLIEscherichia coli K12 cydC26.658.8541ABC transporter
22785778219688321980041122gp: SC9C7_2Streptomyces coelicolor A3(2)35.362.6388hypothetical protein
SC9C7.02
2279577921984472198007441
22805780219847521997581284pir: A72322Thermotoga maritima MSB821.043.7405hypothetical protein
TM0896
22815781219980822010701263sp: HIPO_CAMJECampylobacter jejuni ATCC32.964.3353peptidase
43431 hipO
2282578222014082201073336pir: S38197Arabidopsis thaliana SUC127.151.9133sucrose transport protein
2283578322015842201450135
2284578422018692201594276
22855785220454122019922550prf: 2513410AThermococcus litoralis malP36.167.4814maltodextrin phosphorylase/
glycogen phosphorylase
2286578622054902204591900sp: YFIE_BACSUBacillus subtilis 168 yfiE33.966.4295hypothetical protein
2287578722082492207302948sp: LGT_STAAUStaphylococcus aureus FDA 48531.465.5264prolipoprotein diacylglyceryl
lgttransferase
2288578822091672208367801sp: TRPG_EMENIEmericella nidulans trpC29.662.1169indole-3-glycerol-phosphate
synthase/anthranilate synthase
component II
2289578922098882209232657pir: H70556Mycobacterium tuberculosis29.458.8228hypothetical membrane protein
H37Rv Rv1610
2290579022102732209920354sp: HIS3_RHOSHRhodobacter sphaeroides ATCC52.879.889phosphoribosyl-AMP cyclohydrolase
17023 hisl
2291579122110462210273774sp: HIS6_CORGCorynebacterium glutamicum97.397.7258cyclase
AS019 hisF
2292579222118752211051825prf: 2419176BCorynebacterium glutamicum94.094.0241inositol monophosphate
AS019 impAphosphatase
2293579322126192211882738gp: AF051846_1Corynebacterium glutamicum95.997.6245phosphoribosylformimino-5-
AS019 hisAaminoimidazole carboxamide
ribotide isomerase
2294579422132732212641633gp: AF060558_1Corynebacterium glutamicum86.792.4210glutamine amidotransferase
AS019 hisH
22955795221558622143211266sp: CMLR_STRLIStreptomyces lividans 66 cmlR25.654.0402chloramphenicol resistance protein
or transmembrane transport protein
2296579622158632215639225
2297579722164742215869606sp: HIS7_STRCOStreptomyces coelicolor A3(2)52.581.8198imidazoleglycerol-phosphate
hisBdehydratase
22985798221759122164941098sp: HIS8_STRCOStreptomyces coelicolor A3(2)57.279.3362histidinol-phosphate
hisCaminotransferase
22995799221892522176001326sp: HISX_MYCSMMycobacterium smegmatis63.885.7439histidinol dehydrogenase
ATCC 607 hisD
23005800221915922203581200gp: SPBC215_13Schizosaccharomyces pombe27.254.4342serine-rich secreted protein
SPBC215.13
2301580122211092220459651
2302580222216112221919309
2303580322218282221187642prf: 2321269ALeishmania donovani SAcP-129.459.7211histidine secretory acid phosphatase
2304580422219582222518561pir: RPECR1Escherichia coli plasmid RP128.960.8204tet repressor protein
tetR
23055805222252822250352508prf: 2307203BSulfolobus acidocaldarius treX47.475.5722glycogen debranching enzyme
2306580622251492225949801pir: E70572Mycobacterium tuberculosis50.076.0258hypothetical protein
H37Rv Rv2622
2307580722267632225990774gp: SC2G5_27Streptomyces coelicolor A3(2)29.955.2268oxidoreductase
SC2G5.27c gip
23085808222777922267691011prf: 2503399ASinoRhizobium meliloti ldhA35.060.9343myo-inositol 2-dehydrogenase
2309580922279062228901996sp: GALR_ECOLIEscherichia coli K12 galR30.464.4329galactitol utilization operon repressor
2310581022298962229099798sp: FHUC_BACSUBacillus subtilis 168 fhuC32.968.3246ferrichrome transport ATP-binding
protein or ferrichrome ABC
transporter
23115811223093722299001038prf: 2423441EVibrio cholerae hutC36.871.1332hemin permease
2312581222312942230947348pir: G70046Bacillus subtilis 168 yvrC30.168.0103iron-binding protein
2313581322319322231339594pir: G70046Bacillus subtilis 168 yvrC34.667.6182iron-binding protein
2314581422324562232016441sp: YTFH_ECOLIEscherichia coli K12 ytfH38.173.5113hypothetical protein
23155815223292822340701143gp: SCI8_12Streptomyces coelicolor A3(2)23.450.1355DNA polymerase III epsilon chain
SCI8.12
2316581622341582234763606
23175817223485222372842433pir: S65769Arthrobacter sp. Q36 treY42.068.6814maltooligosyl trehalose synthase
23185818223733122383531023gp: AE002006_4Deinococcus radiodurans27.652.8322hypothetical protein
DR1631
2319581922390922238694399
2320582022400422239845198
2321582122402462240058189
23225822224056322395081056
23235823224068122417241044sp: LXA1_PHOLUPhotorhabdus luminescens20.554.4375alkanal monooxygenase alpha chain
ATCC 29999 luxA
2324582422421152241738378gp: SC7H2_5Streptomyces coelicolor A3(2)58.379.2120hypothetical protein
SC7H2.05
2325582522423592242129231
23265826224303522448191785pir: S65770Arthrobacter sp. Q36 treZ46.372.4568maltooligosyltrehalose
trehalohydrolase
2327582722430432242393651sp: YVYE_BACSUBacillus subtilis 16836.572.4214hypothetical protein
23285828224617122448641308sp: THD1_CORGLCorynebacterium glutamicum99.399.3436threonine dehydratase
ATCC 13032 ilvA
2329582922463862246892507
2330583022464502246295156
23315831224820822470061203pir: S57636Catharanthus roseus metE22.749.6415Corynebacterium glutamicum AS019
23325832225193922483583582prf: 2508371AStreptomyces coelicolor A3(2)53.380.51183DNA polymerase III
dnaE
2333583322520172252856840sp: RARD_ECOLIEscherichia coli K12 rarD37.673.8279chloramphenicol sensitive protein
2334583422531922253659468sp: HISJ_CAMJECampylobacter jejuni DZ72 hisJ21.555.7149histidine-binding protein precursor
2335583522537252254642918pir: D69548Archaeoglobus fulgidus AF238822.764.7198hypothetical membrane protein
2336583622555582254683876sp: GS39_BACSUBacillus subtilis 168 ydaD48.280.0280short chain dehydrogenase or
general stress protein
23375837225702422557381287sp: DCDA_PSEAEPseudomonas aeruginosa lysA22.947.6445diaminopimelate (DAP)
decarboxylase
2338583822593122258362951sp: CYSM_ALCEUAlcaligenes eutrophus CH3432.864.3314cysteine synthase
cysM
2339583922599992259421579
2340584022609312260002930sp: RLUD_ECOLIEscherichia coli K12 rluD36.561.0326ribosomal large subunit
pseudouridine synthase D
2341584122614672260934534sp: LSPA_PSEFLPseudomonas fluorescens NCIB33.861.7154lipoprotein signal peptidase
10586 lspA
23425842226168822626891002
23435843226285022644991650pir: S67863Streptomyces antibioticus oleB36.464.0550oleandomycin resistance protein
2344584422649962265298303
2345584522651082264509600prf: 2422382PRhodococcus erythropolis orf1736.757.6158hypothetical protein
2346584622654202266394975sp: ASPG_BACLIBacillus licheniformis31.262.0321L-asparaginase
23475847226829722668971401sp: DINP_ECOLIEscherichia coli K12 dinP31.860.7371DNA-damage-inducible protein P
2348584822692452268388858sp: YBIF_ECOLIEscherichia coli K12 ybiF31.561.5286hypothetical membrane protein
23495849227026122692601002gp: SCF51_6Streptomyces coelicolor A3(2)44.373.1334transcriptional regulator
SCF51.06
2350585022703042270435132
2351585122708842270258627gp: SCF51_5Streptomyces coelicolor A3(2)42.067.0212hypothetical protein
SCF51.05
23525852227414922709883162sp: SYIC_YEASTSaccharomyces cerevisiae38.565.41066isoleucyl-tRNA synthetase
A364A YBL076C ILS1
2353585322746882274473216
23545854227586122747671095
2355585522766372276353285pir: F70578Mycobacterium tuberculosis46.373.282hypothetical membrane protein
H37Rv Rv2146c
2356585622773362276881456gp: BLFTSZ_6Brevibacterium lactofermentum99.399.3152hypothetical protein (putative YAK 1
orf6protein)
2357585722780782277416663sp: YFZ1_CORGLCorynebacterium glutamicum97.799.6221hypothetical protein
2358585822788592278122738prf: 2420425CBrevibacterium lactofermentum99.2100.0246hypothetical protein
yfih
2359585922791552279640486GP: AB028868_1Mus musculus P4(21)n39.051.0117hypothetical protein
23605860228021522788901326sp: FTSZ_BRELABrevibacterium lactofermentum98.698.6442cell division protein
ftsZ
2361586122811352280470666gsp: W70502Corynebacterium glutamicum99.6100.0222cell division initiation protein or cell
ftsQdivision protein
23625862228262322811661458gp: AB015023_1Corynebacterium glutamicum99.499.8486UDP-N-acetylmuramate—alanine
murCligase
23635863228377622826611116gp: BLA242646_3Brevibacterium lactofermentum98.999.5372UDP-N-acetylglucosamine-N-
ATCC 13869 murGacetylmuramyl-(pentapeptide)
pyrophosphoryl-undecaprenol N-
acetylglucosamine pyrophosphoryl-
undecaprenol N-acetylglucosamine
23645864228543122837821650gp: BLA242646_3Brevibacterium lactofermentum99.499.6490cell division protein
ATCC 13869 ftsW
2365586522859042285437468gp: BLA242646_1Brevibacterium lactofermentum99.199.1110UDP-N-acetylmuramoylalanine-D-
ATCC 13869 murDglutamate ligase
2366586622862722286655384
2367586722864992286831333
23685868228795922868621098sp: MRAY_ECOLIEscherichia coli K12 mraY38.663.8365phospho-n-acetylmuramoyl-
pentapeptide
23695869228951022879691542sp: MURF_ECOLIEscherichia coli K12 murF35.064.2494UDP-N-acetylmuramoylalanyl-D-
glutamyl-2,6-diaminoplmelate-D-
alanyl-D-alanyl ligase
23705870229107322895231551sp: MURE_BACSUBacillus subtilis 168 murE37.767.6491UDP-N-acetylmuramoylalanyl-D-
glutamyl-2,6-diaminopimelate-D-
alanyl-D-alanyl ligase
2371587122911972290973225GSP: Y33117Brevibacterium lactofermentum100.0100.057penicillin binding protein
ORF2 pbp
23725872229316422912121953pir: S54872Pseudomonas aeruginosa pbpB28.258.8650penicillin-binding protein
2373587322941172293323795
23745874229512722941171011pir: A70581Mycobacterium tuberculosis55.179.3323hypothetical protein
H37Rv Rv2165c
2375587522958042295376429gp: MLCB268_11Mycobacterium leprae72.088.8143hypothetical membrane protein
MLCB268.11c
2376587622968982296512387pir: C70935Mycobacterium tuberculosis39.469.3137hypothetical protein
H37Rv Rv2169c
2377587722976532297231423
2378587822978662298438573gp: MLCB268_13Mycobacterium leprae36.365.3190hypothetical protein
MLCB268.13
2379587922994282298451978sp: METF_STRLIStreptomyces lividans 132642.670.63035,10-methylenetetrahydrofolate
metFreductase
23805880229952423006361113pir: S32168Myxococcus xanthus DK105030.162.0329dimethylallyltranstransferase
ORF1
23815881230070623021751470gp: MLCB268_16Mycobacterium leprae35.769.6484hypothetical membrane protein
MLCB268.17
2382588223021792302685507
2383588323026192302251369pir: A70936Mycobacterium tuberculosis43.268.8125hypothetical protein
H37Rv Rv2175c
23845884230283323049802148gp: AB019394_1Streptomyces coelicolor A3(2)34.262.4684eukaryotic-type protain kinase
pkaF
2385588523036902303040651
23865886230498323062181236gp: MLCB268_21Mycobacterium leprae30.758.4411hypothetical membrane protein
MLCB268.23
23875887230631423076211308pir: G70936Mycobacterium tuberculosis30.462.0434hypothetical membrane protein
H37Rv Rv2181
23885888230908223076971386gp: AF260581_2Amycolatopsis mediterranei66.987.94623-deoxy-D-arabino-heptulosonate-7-
phosphate synthase
2389588923096762309173504gp: MLCB268_20Mycobacterium leprae58.477.7166hypothetical protein
MLCB268.21c
23905890230983523122522418pir: G70936Mycobacterium tuberculosis35.164.5428hypothetical membrane protein
H37Rv Rv2181
23915891231236023138081449sp: CSP1_CORGLCorynebacterium glutamicum28.257.1440major secreted protein PS1 protein
(Brevibacterium flavum) ATCCprecursor
17965 csp1
2392589223138332314036204
2393589323140922313916177
23945894231542323142361188gp: AF096280_3Corynebacterium glutamicum100.0100.0249hypothetical membrane protein
ATCC 13032
2395589523164122315678735gp: AF096280_2Corynebacterium glutamicum100.0100.0245acyltransferase
ATCC 13032
23965896231877523176331143gp: SC6G10_5Streptomyces coelicolor A3(2)50.175.7383glycosyl transferase
SC6G10.05c
23975897231985023188041047sp: P60_LISIVListeria ivanovii iap26.460.8296protein P60 precursor (invasion-
associated-protein)
2398589823205942319968627sp: P60_LISGRListeria grayi iap33.061.3191protein P60 precursor (invasion-
associated-protein)
23995899232307323214721602prf: 2503462KHeliobacillus mobilis petB34.364.7201ubiquinol-cytochrome c reductase
cytochrome b subunit
2400590023237592323088672gp: AF107888_1Streptomyces lividans qcrA37.957.1203ubiquinol-cytochrome c reductase
iron-sulfur subunit (Rieske [eFe-2S]
iron-sulfur protein cyoB
2401590123251952324311885sp: Y005_MYCTUMycobacterium tuberculosis58.683.1278ubiquinol-cytochrome c reductase
H37Rv Rv2194 qcrCcytochrome c
2402590223258872325273615sp: COX3_SYNVUSynechococcus vulcanus36.770.7188cytochrome c oxidase subunit III
2403590323262732326121153
2404590423269002326472429sp: Y00A_MYCTUMycobacterium tuberculosis38.671.0145hypothetical membrane protein
H37Rv Rv2199c
24055905232799723269211077sp: COX2_RHOSHRhodobacter sphaeroides ctaC28.753.9317cytochrome c oxidase subunit II
24065906232851623304351920gp: AB029550_1Corynebacterium glutamicum99.799.8640glutamine-dependent
KY9611 ltsAamidotransferase or asparagine
synthetase (lysozyme insensitivity
protein)
2407590723309272330586342gp: AB029550_2Corynebacterium glutamicum100.0100.0114hypothetical protein
KY9611 orf1
2408590823312002331967768gp: MLCB22_2Mycobacterium leprae35.060.2246hypothetical membrane protein
MLCB22.07
2409590923319742332495522pir: S52220Rhodobacter capsulatus cobP43.064.0172cobinamide kinase
24105910233251223336001089sp: COBU_PSEDEPseudomonas denitrificans37.866.9341nicotinate-nucleotide—
cobUdimethylbenzimidazole
phosphoribosyltransferase
2411591123336152334535921sp: COBV_PSEDEPseudomonas denitrificans cobV25.349.8305cobalamin (5′-phosphate) synthase
2412591223347172334481237
2413591323357412335028714prf: 2414335AStreptomyces clavuligerus car38.668.5241clavulanate-9-aldehyde reductase
24145914233705123359151137sp: ILVE_MYCTUMus musculus BCAT140.170.3364branched-chain amino acid
aminotransferase
24155915233723523387341500gp: PPU010261_1Pseudomonas putida ATCC36.365.9493leucyl aminopeptidase
12633 pepA
2416591623391402338748393prf: 2110282ASaccharopolyspora erythraea40.267.097hypothetical protein
ORF1
24175917233926923412932025gp: AF047034_2Streptomyces seoulensis pdhB48.968.5691dihydrolipoamide acetyltransferase
24185918234080423394401365
2419591923414122342164753gp: AB020975_1Arabidopsis thaliana36.765.7210lipoyltransferase
24205920234230423433471044sp: LIPA_PELCAPelobacter carbinolicus GRA BD144.670.9285lipoic acid synthetase
lipA
2421592123434792344258780sp: Y00U_MYCTUMycobacterium tuberculosis45.576.7257hypothetical membrane protein
H37Rv Rv2219
24225922234443123460471617sp: YIDE_ECOLIEscherichia coli K12 yidE32.967.8559hypothetical membrane protein
24235923234749123462891203gp: AF189147_1Corynebacterium glutamicum100.0100.0401transposase (ISCg2)
ATCC 13032 tnp
2424592423475052347804300
2425592523485482348078471gp: SC5F7_34Streptomyces coelicolor A3(2)41.463.7157hypothetical membrane protein
SC5F7.04c
2426592623506202350408213
242759272351022235199697531.044.0145mutator mutT domain protein
2428592823513102350912399pir: B72308Thermotoga maritima MSB836.765.6128hypothetical protein
TM1010
2429592923519092351310600
2430593023519802352828849sp: LUXA_VIBHAVibrio harveyi luxA25.060.9220alkanal monooxygenase alpha chain
(bacterial luciferase alpha chain)
2431593123528332353225393pir: A72404Thermotoga maritima MSB840.573.0111protein synthesis inhibitor
TM0215(translation initiation inhibitor)
2432593223551562355398243
2433593323554402355180261
24345934235552123568431323prf: 2203345HEscherichia coli hpaX21.953.44334-hydroxyphenylacetate permease
2435593523567942357354561gp: SCGD3_10Streptomyces coelicolor A3(2)42.472.8158transmembrane transport protein
SCGD3.10c
2436593623572642357707444gp: SCGD3_10Streptomyces coelicolor A3(2)31.466.1118transmembrane transport protein
SCGD3.10c
2437593723574842357290195
2438593823577262358130405
2439593923586952358153543
2440594023594162358772645sp: HMUO_CORDICorynebacterium diphtheriae C757.978.0214heme oxygenase
hmuO
24415941236274823596143135gp: SCY17736_4Streptomyces coelicolor A3(2)43.467.0809glutamate-ammonia-ligase
glnEadenylyltransferase
24425942236415523628181338sp: GLNA_THEMAThermotoga maritima MSB843.573.0441glutamine synthetase
glnA
24435943236435223654551104gp: SCE9_39Streptomyces coelicolor A3(2)26.854.1392hypothetical protein
SCE9.39c
24445944236558723674131827sp: Y017_MYCTUMycobacterium tuberculosis33.458.2601hypothetical protein
H37Rv Rv2226
2445594523676522367473180gp: SCC75A_11Streptomyces coelicolor A3(2)38.955.654hypothetical protein
SCC75A.11c.
24465946236779123690831293sp: GAL1_HUMANHomo sapiens galK124.953.7374galactokinase
24475947237038123691161266gp: AF174645_1Brucella abortus vacB27.154.5358virulence-associated protein
2448594823704232370908486
24495949237255723714121146sp: Y019_MYCTUMycobacterium tuberculosis54.775.1382bifunctional protein (ribonuclease H
H37Rv Rv2228cand phosphoglycerate mutase)
2450595023725612373289729
2451595123732892372573717sp: Y01A_MYCTUMycobacterium tuberculosis26.558.6249hypothetical protein
H37Rv Rv2229c
24525952237446223733231140sp: Y01B_MYCTUMycobacterium tuberculosis49.276.2378hypothetical protein
H37Rv Rv2230c
2453595323745442375197654sp: GPH_ECOLIEscherichia coli K12 gph26.054.4204phosphoglycolate phosphatase
2454595423752142375684471sp: PTPA_STRCOStreptomyces coelicolor A3(2)46.263.5156low molecular weight protein-
SCQ11.04c ptpAtyrosine-phosphatase
2455595523757672376720954sp: Y01G_MYCTUMycobacterium tuberculosis40.965.5281hypothetical protein
H37Rv Rv2235
2456595623773902376998393sp: YI21_BURCEBurkholderia cepacia32.656.6129insertion element (IS402)
2457595723777262377484243
2458595823778992378276378gp: SC8F4_22Streptomyces coelicolor A3(2)30.457.8135transcriptional regulator
SC8F4.22c
2459595923782922378489198
2460596023793122378884429sp: Y01K_MYCTUMycobacterium tuberculosis55.277.6134hypothetical protein
H37Rv Rv2239c
2461596123794262379770345
24625962238003323827442712gp: AF047034_4Streptomyces seoulensis pdhA55.978.9910pyruvate dehydrogenase component
24635963238224023807651476
2464596423836152382827789sp: GLNQ_ECOLIEscherichia coli K12 glnQ33.762.8261ABC transporter or glutamine
transport ATP-binding protein
2465596523844642385426963
2466596623845092383622888sp: RBSC_BACSUBacillus subtilis 168 rbsC25.458.7283ribose transport system permease
protein
2467596723854472384509939pir: H71693Rickettsia prowazekii Madrid E26.262.9286hypothetical protein
RP367
2468596823857712386580810sp: CBPA_DICDIDictyostelium discoideum AX241.655.2125calcium binding protein
cbpA
2469596923862842385913372
24705970238762723866141014gp: SC6G4_24Streptomyces coelicolor A3(2)29.655.7352lipase or hydrolase
SC6G4.24
2471597123876672387957291sp: ACP_MYXXAMyxococcus xanthus ATCC42.780.075acyl carier protein
25232 acpP
2472597223879972388821825sp: NAGD_ECOLIEscherichia coli K12 nagD43.975.5253N-acetylglucosamine-6-phosphate
deacetylase
24735973238883823898691032gp: AE001968_4Deinococcus radiodurans33.665.7289hypothetical protein
DR1192
2474597423909042390434471
2475597523920082391184825gp: SC4A7_8Streptomyces coelicolor A3(2)52.475.3271hypothetical protein
SC4A7.08
2476597623925662392075492
2477597723933492392579771
2478597823934252393970546
2479597923944372393973465
2480598023945942394935342
24815981239520423967631560sp: PPBD_BACSUBacillus subtilis 168 phoD34.264.7530alkaline phosphatase D precursor
2482598223959862395273714
24835983239726423990991836gp: SCI51_17Streptomyces coelicolor A3(2)44.473.1594hypothetical protein
SCI51.17
2484598423991582399397240pir: G70661Mycobacterium tuberculosis41.272.168hypothetical protein
H37Rv Rv2342
2485598524003422399668675
24865986240130323994051899prf: 2413330BMycobacterium smegmatis59.182.9633DNA primase
dnaG
2487598724013732401834462gp: XXU39467_1Streptomyces aureofaciens BMK49.067.498ribonuclease Sa
2488598824018382402080243
2489598924031652402530636
24905990240401224021441869gp: AF058788_1Mycobacterium smegmatis59.182.2636L-glutamine: D-fructose-6-phosphate
mc2155 glmSamidotransferase
2491599124045232404846324
24925992240567124068221152
24935993240625824049871272prf: 2413330AMycobacterium smegmatis dgt54.676.3414deoxyguanosinetriphosphate
triphosphohydrolase
2494599424069362406262675gp: NMA1Z2491_235Neisseria meningitidis NMA025130.459.7171hypothetical protein
24955995240699324090292037pir: B70662Mycobacterium tuberculosis31.163.6692hypothetical protein
H37Rv Rv2345
2496599624102642409779486gp: AE003565_26Drosophila melanogaster24.654.4138hypothetical protein
CG10592
2497599724108612410280582
24985998241233824109561383pir: S58522Thermus aquaticus HB846.169.9508glycyl-tRNA synthetase
2499599924125802412948369pir: E70585Mycobacterium tuberculosis49.473.089bacterial regulatory protein, arsR
H37Rv Rv2358 furBfamily
2500600024129922413423432sp: FUR_ECOLIEscherichia coli K12 fur34.970.5132ferric uptake regulation protein
25016001241356824151181551pir: A70539Mycobacterium tuberculosis24.846.7529hypothetical protein (conserved in
H37Rv Rv1128cC. glutamicum?)
2502600224160892415298792gp: AF162938_1Streptomyces coelicolor A3(2)40.667.0224hypothetical membrane protein
h3u
2503600324170992416371729sp: UPPS_MICLUMicrococcus luteus B-P 26 uppS43.471.2233undecaprenyl diphosphate synthase
2504600424179472417222726pir: A70586Mycobacterium tuberculosis45.774.3245hypothetical protein
H37Rv Rv2362c
2505600524188832417969915gp: AF072811_1Streptococcus pneumoniae era39.570.3296Era-like GTP-binding protein
25066006242030924189901320sp: Y1DE_MYCTUMycobacterium tuberculosis52.882.4432hypothetical membrane protein
H37Rv Rv2366
2507600724209002420313588sp: YN67_MYCTUMycobacterium tuberculosis65.086.0157hypothetical protein
H37Rv Rv2367c
2508600824209732421236264GSP: Y75650Neisseria meningitidis45.050.085Neisserial polypeptides predicted to
be useful antigens for vaccines and
diagnostics
25096009242194924209001050sp: PHOL_MYCTUMycobacterium tuberculosis61.184.6344phosphate starvation inducible
H37Rv Rv2368c phoHprotein
2510601024226972421975723gp: SCC77_19Streptomyces coelicolor A3(2)44.075.4248hypothetical protein
SCC77.19c.
2511601124228502423791942
25126012242384524227001146prf: 2421342BStreptomyces albus dnaJ247.177.4380heat shock protein dnaJ
25136013242493724239151023prf: 2421342AStreptomyces albus hrcA48.279.6334heat-inducible transcriptional
repressor (groEL repressor)
2514601424259542424965990prf: 2318256ABacillus stearothermophilus33.164.1320oxygen-independent
hemNcoproporphyrinogen III oxidase
2515601524261812426699519sp: AGA1_YEASTSaccharomyces cerevisiae36.664.9134agglutinin attachment subunit
YNR044W AGA1precursor
2516601624274682426776693
2517601724281842427807378
25186018243002824281841845gp: SC6G10_4Streptomyces coelicolor A3(2)48.075.1611long-chain-fatty-acid—CoA ligase
SC6G10.04
25196019243029624324132118sp: MALQ_ECOLIEscherichia coli K12 malQ28.355.47384-alpha-glucanotransferase
25206020243250824343701863gp: AB005752_1Lactobacillus brevis plasmid29.564.4604ABC transporter, Hop-Resistance
horAprotein
2521602124338682433614255GSP: Y74827Neisseria gonorrhoeae44.051.068Neisserial polypeptides predicted to
be useful antigens for vaccines and
diagnostics
2522602224342072433875333GSP: Y74829Neisseria meningitidis47.053.0107polypeptides predicted to be useful
antigens for vaccines and
diagnostics
2523602324346192434440180
2524602424347762434573204
25256025243683824348052034sp: DCP_SALTYSalmonella typhimurium dcp40.368.3690peptidyl-dipeptidase
25266026243687124380491179gp: AF064523_1Anisopteromalus calandrae24.145.7453carboxylesterase
25276027243811324399061794pir: G70983Mycobacterium tuberculosis65.284.9594glycosyl hydrolase or trehalose
H37Rv Rv0126synthase
25286028243990624409941089pir: H70983Mycobacterium tuberculosis32.158.8449hypothetical protein
H37Rv Rv0127
2529602924415892441005585pir: T07979Chlamydomonas reinhardtii ipi131.857.7189isopentenyl-diphosphate Delta-
isomerase
2530603024416692441890222
2531603124423552442792438
25326032244335624416021755
2533603324440152443356660
2534603424445512444033519
2535603524447352445709975gp: CORCSLYS_1Corynebacterium glutamicum99.4100.0325beta C-S lyase (degradation of
ATCC 13032 aecDaminoethylcysteine)
25366036244571624469931278sp: BRNQ_CORGLCorynebacterium glutamicum99.8100.0426branched-chain amino acid transport
ATCC 13032 brnQsystem carrier protein (isoleucine
uptake)
2537603724470212447998978sp: LUXA_VIBHAVibrio harveyi luxA21.649.0343alkanal monooxygenase alpha chain
2538603824508442450323522
2539603924517852450859927gp: AF155772_2SinoRhizobium meliloti mdcF25.960.5324malonate transporter
25406040245463724517942844sp: GLCD_ECOLIEscherichia coli K12 glcD27.755.1483glycolate oxidase subunit
2541604124547252455435711sp: YDFH_ECOLIEscherichia coli K12 ydfH25.665.0203transcriptional regulator
2542604224557332455452282
25436043245706624557201347sp: YGIK_SALTYSalmonella typhimurium ygiK22.557.6467hypothetical protein
2544604424577592457337423
25456045245786324593711509sp: HBPA_HAEINHaemophilus influenzae Rd27.555.5546heme-binding protein A precursor
HI0853 hbpA(hemin-binding lipoprotein)
2546604624593712460336966sp: APPB_BACSUBacillus subtilis 168 appB40.073.3315oligopeptide ABC transporter
(permease)
2547604724603402461167828sp: DPPC_ECOLIEscherichia coli K12 dppC43.274.5271dipeptide transport system
permease protein
25486048246116324625991437prf: 2306258MREscherichia coli K12 oppD37.466.4372oligopeptide transport ATP-binding
protein
2549604924620492461543507PIR: G72536Aeropyrum pernix K1 APE158035.044.0106hypothetical protein
2550605024631502462602549pir: D70367Aquifex aeolicus VF5 aq_76829.358.0157hypothetical protein
2551605124632412464143903prf: 2514301ARhizobium etli rbsK41.065.0300ribose kinase
25526052246434424657681425gp: SCM2_16Streptomyces coelicolor A3(2)39.964.6466hypothetical membrane protein
SCM2.16c
2553605324657672465465303
2554605424670092466038972sp: NTCI_HUMANHomo sapiens31.361.6284sodium-dependent transporter or
odium Bile acid symporter family
2555605524670772467922846gp: AF195243_1Chlamydomonas reinhardtii28.551.2295apospory-associated protein C
2556605624703132470678366
2557605724722502472819570sp: THIX_CORGLCorynebacterium glutamicum100.0100.0133thiamine biosynthesis protein x
ATCC 13032 thiX
2558605824734802472893588sp: VG66_BPMDMycobacteriophage D29 6642.665.5197hypothetical protein
25596059247365324755421890sp: BETP_CORGLCorynebacterium glutamicum39.871.7601glycine betaine transporter
ATCC 13032 betP
2560606024764972477492996
25616061247764424792511608
2562606224793792479762384
25636063248120824798981311prf: 2320266CRhodobacter capsulatus dctM34.671.9448large integral C4-dicarboxylate
membrane transport protein
2564606424816922481213480gp: AF186091_1Klebsiella pneumoniae dctQ33.973.7118small integral C4-dicarboxylate
membrane transport protein
2565606524824802481734747sp: DCTP_RHOCARhodobacter capsulatus B1028.259.0227C4-dicarboxylate-binding
dctPperiplasmic protein precursor
2566606624838452484087243PRF: 1806416ALycopersicon esculentum63.073.046extensin I
(tomato)
25676067248439224825481845sp: LEPA_BACSUBacillus subtilis 168 lepA58.783.6603GTP-binding protein
2568606824846612485269609pir: H70683Mycobacterium tuberculosis41.669.7185hypothetical protein
H37Rv Rv2405
2569606924854732485733261sp: RS20_ECOLIEscherichia coli K12 rpsT48.272.98530S ribosomal protein S20
2570607024864692485801669sp: RHTC_ECOLIEscherichia coli K12 rhtC30.067.1210thrreonine efflux protein
2571607124868812486477405gp: SC6D7_25Streptomyces coelicolor A3(2)61.280.6129ankyrin-like protein
SC6D7.25.
2572607224878842486910975pir: H70684Mycobacterium tuberculosis46.074.1313hypothetical protein
H37Rv Rv2413c
25736073248945024879121539sp: CME3_BACSUBacillus subtilis 168 comEC21.449.7527late competence operon required for
DNA binding and uptake
2574607424901542489573582sp: CME1_BACSUBacillus subtilis 168 comEA30.863.6195late competence operon required for
DNA binding and uptake
2575607524909112491732822
2576607624911112490290822gp: SCC123_7Streptomyces coelicolor A3(2)34.866.3273hypothetical protein
SCC123.07c.
2577607724918582491151708pir: F70685Mycobacterium tuberculosis46.866.4235phosphoglycerate mutase
H37Rv Rv2419c
2578607824923432491873471pir: G70685Mycobacterium tuberculosis55.686.3117hypothetical protein
H37Rv Rv2420c
2579607924931782492501678gp: SCC123_17Streptomyces coelicolor A3(2)68.085.3197hypothetical protein
SCC123.17c.
25806080249423724932151023
25816081249563424943391296sp: PROA_CORGLCorynebacterium glutamicum99.199.8432gamma-glutamyl phosphate
ATCC 17965 proAreductase or glutamate-5-
semialdehyde dehydrogenase
2582608224966072495696912sp: YPRA_CORGLCorynebacterium glutamicum99.3100.0304D-isomer specific 2-hydroxyacid
ATCC 17965 unkdhdehydrogenase
2583608324968032497513711
25846084249951124980091503gp: D87915_1Streptomyces coelicolor A3(2)58.978.2487GTP-binding protein
obg
25856085249978325016691887sp: PBUX_BACSUBacillus subtilis 168 pbuX39.177.3422xanthine permease
2586608625025772501735843pir: I40838Corynebacterium sp. ATCC61.281.92762,5-diketo-D-gluconic acid reductase
31090
2587608725027352503355621
2588608825038702504265396
2589608925042472503984264sp: RL27_STRGRStreptomyces griseus IFO1318980.392.68150S ribosomal protein L27
rpmA
2590609025046022504300303prf: 2304263AStreptomyces griseus IFO1318956.482.210150S ribosomal protein L21
obg
25916091250709825048312268sp: RNE_ECOLIEscherichia coli K12 rne30.156.6886ribonuclease E
2592609225071152507663549
2593609325071382507710573
2594609425080942508840747
2595609525089222509530609gp: SCF76_8Streptomyces coelicolor A3(2)61.082.6195hypothetical protein
SCF76.08c
25966096251083025095231308plr: S43613Corynebacterium glutamicum99.1100.0436transposase (insertion sequence
ATCC 31831IS31831)
2597609725110462511423378gp: SCF76_8Streptomyces coelicolor A3(2)51.376.9117hypothetical protein
SCF76.08c.
2598609825114272511876450gp: SCF76_9Streptomyces coelicolor A3(2)37.867.8143hypothetical protein
SCF76.09
2599609925123562511949408gp: AF069544_1Mycobacterium smegmatis ndk70.989.6134nucleoside diphosphate kinase
2600610025127682512409360
2601610125128032513144342gp: AE002024_10Deinococcus radiodurans R134.867.492hypothetical protein
DR1844
2602610225136182513154465pir: H70515Mycobacterium tuberculosis36.664.3112hypothetical protein
H37Rv Rv1883c
2603610325141142513692423pir: E70863Mycobacterium tuberculosis33.968.6118hypothetical protein
H37Rv Rv2446c
26046104251548725141141374prf: 2410252BStreptomyces coelicolor A3(2)55.479.6451folyl-polyglutamate synthetase
folC
2605610525156622516273612
2606610625162432516956714
2607610725170892517751663
26086108251833625156372700sp: SYV_BACSUBacillus subtilis 168 balS45.572.1915valyl-tRNA synthetase
26096109251997225183981575pir: A38447Bacillus subtilis 168 oppA24.258.5521oligopeptide ABC transport system
substrate-binding protein
26106110252020925216601452sp: DNAK_BACSUBacillus subtilis 168 dnaK26.254.9508heat shock protein dnaK
2611611125222512521667585gp: ECU89166_1Eikenella corrodens ATCC42.971.2170lysine decarboxylase
23824
2612611225232482522265984sp: MDH_THEFLThermus aquaticus ATCC 3392356.476.5319malate dehydrogenase
mdh
2613611325235612524337777gp: SC4A10_33Streptomyces coelicolor A3(2)24.656.5207transcriptional regulator
SC4A10.33
2614611425249152524340576gp: AF065442_1Vibrio cholerae aphA26.051.4208hypothetical protein
26156115252509925262261128prf: 2513416FAcinetobacter sp. vanA39.568.6357vanillate demethylase (oxygenase)
2616611625262332527207975gp: FSU12290_2Sphingomonas flava ATCC32.859.2338pentachlorophenol 4-
39723 pcpDmonooxygenase reductase
26176117252713525285591425prf: 2513416GAcinetobacter sp. vanK40.876.8444transport protein
2618611825294802528551930gp: KPU95087_7Klebsiella pneumoniae mdcF28.058.4286malonate transporter
26196119253076125294841278prf: 2303274ABacillus subtilis clpX59.885.8430class-III heat-shock protein or ATP-
dependent protease
26206120253089125319761086gp: SCF55_28Streptomyces coelicolor A3(2)45.673.0366hypothetical protein
SCF55.28c
2621612125326012531969633gp: AF109386_2Streptomyces sp. 2065 pcaJ63.385.7210succinyl CoA: 3-oxoadipate CoA
transferase beta subunit
2622612225333532532604750gp: AF109386_1Streptomyces sp. 2065 pcal60.284.5251succinyl CoA: 3-oxoadipate CoA
transferase alpha subunit
2623612325333912534182792prf: 2408324FRhodococcus opacus 1CP pcaR58.282.5251protocatechuate catabolic protein
26246124253420125354241224prf: 2411305DRalstonia eutropha bktB44.871.9406beta-ketothiolase
2625612525351682534257912
2626612625354302536182753prf: 2408324ERhodococcus opacus pcaL50.876.62563-oxoadipate enol-lactone hydrolase
and 4-carboxymuconolactone
decarboxylase
26276127253619625382562061gp: SCM1_10Streptomyces coelicolor A3(2)23.643.0825transcriptional regulator
SCM1.10
2628612825386132538248366prf: 2408324ERhodococcus opacus pcaL78.389.61153-oxoadipate enol-lactone hydrolase
and 4-carboxymuconolactone
decarboxylase
2629612925395532540230678
26306130253973125386161116prf: 2408324DRhodococcus opacus pcaB39.863.44373-carboxy-cis, cis-muconate
cycloisomerase
2631613125403202539709612prf: 2408324CRhodococcus opacus pcaG49.570.6214protocatechuate dioxygenase alpha
subunit
2632613225410242540335690prf: 2408324BRhodococcus opacus pcaH74.791.2217protocatechuate dioxygenase beta
subunit
26336133254235025411871164pir: G70506Mycobacterium tuberculosis26.448.7273hypothetical protein
H37Rv Rv0336
2634613425428022542512291prf: 2515333BMycobacterium tuberculosis54.481.592muconolactone isomerase
catC
2635613525430432543813771
26366136254393625428181119sp: CATB_RHOOPRhodococcus opacus 1CP catB60.884.7372muconate cycloisomerase
2637613725442622544867606
2638613825448762544022855prf: 2503218ARhodococcus rhodochrous catA72.388.4285catechol 1,2-dioxygenase
2639613925450682544928141
26406140254531525467841470gp: AF134348_1Pseudomonas putida plasmid62.285.6437toluate 1,2 dioxygenase subunit
pDK1 xylX
2641614125468272547318492gp: AF134348_2Pseudomonas putida plasmid60.383.2161toluate 1,2 dioxygenase subunit
pDK1 xylY
26426142254733325488681536gp: AF134348_3Pseudomonas putida plasmid51.581.0342toluate 1,2 dioxygenase subunit
pDK1 xylZ
2643614325488682549695828gp: AF134348_4Pseudomonas putida plasmid30.761.42771,2-dihydroxycyclohexa-3,5-diene
pDK1 xylLcarboxylate dehydrogenase
26446144254977125524552685gp: REU95170_1Rhodococcus erythropolis thcG23.348.6979regulator of LuxR family with ATP-
binding site
26456145255256325539421380sp: PCAK_ACICAAcinetobacter calcoaceticus31.364.4435transmembrane transport protein or
pcaK4-hydroxybenzoate transporter
26466146255402625552671242sp: BENE_ACICAAcinetobacter calcoaceticus29.966.2388benzoate membrane transport
benEprotein
2647614725559402555317624gp: AF071885_2Streptomyces coelicolor M14569.588.3197ATP-dependent Clp protease
clpP2proteolytic subunit 2
2648614825565802555978603gp: AF071885_1Streptomyces coelicolor M14562.185.9198ATP-dependent Clp protease
clpP1proteolytic subunit 1
2649614925565992556748150gp: SIS243537_4Sulfolobus islandicus ORF15442.971.442hypothetical protein
26506150255810625567601347sp: TIG_BACSUBacillus subtilis 168 tig32.166.4417trigger factor (prolyl isomerase)
(chaperone protein)
2651615125586092559103495gp: SCD25_17Streptomyces coelicolor A3(2)32.563.1160hypothetical protein
SCD25.17
2652615225591572560131975sp: PBP4_NOCLANocardia lactamdurans LC41125.350.9336penicillin-binding protein
pbp
2653615325601312560586456prf: 2301342AMus musculus Moa127.858.3115hypothetical protein
2654615425611152561363249
2655615525619202561483438prf: 2513302CCorynebacterium striatum ORF154.273.2142transposase
2656615625620932562242150
2657615725621152561990126prf: 2513302CCorynebacterium striatum ORF157.182.935hypothetical protein
2658615825623412562078264prf: 2513302CCorynebacterium striatum ORF150.778.775transposase
2659615925627762562387390
2660616025629632563847885
2661616125644022563932471sp: LACB_STAAUStaphylococcus aureus NCTC40.071.4140galactose-6-phosphate isomerase
8325-4 lacB
2662616225652452564550696sp: YAMY_BACADBacillus acidopullulyticus ORF226.258.1248hypothetical protein
2663616325662312565623609pir: A70866Mycobacterium tuberculosis56.880.9199hypothetical protein
H37Rv Rv2466c
26646164256634525689452601sp: AMPN_STRLIStreptomyces lividans pepN47.570.5890aminopeptidase N
26656165256921125702931083pir: B70206Borrelia burgdorferi BB085225.158.1358hypothetical protein
26666166257146025703091152
2667616725715102572175666
2668616825721932572348156
2669616925726772572351327gp: AF139916_3Brevibacterium linens ATCC61.581.7104phytoene desaturase
9175 crtI
2670617025729772572807171
2671617125737702573393378
26726172257386425726591206sp: CRTJ_MYXXAMyxococcus xanthus DK105031.263.8381phytoene dehydrogenase
carA2
2673617325747182573843876sp: CRTB_STRGRStreptomyces griseus JA393331.458.6290phytoene synthase
crtB
26746174257589825747801119gp: LMAJ9627_3Listeria monocytogenes lltB25.847.7392multidrug resistance transporter
26756175257721325759811233
26766176257887225772321641gp: SYOATPBP_2Synechococcus elongatus41.371.6538ABC transporter ATP-binding protein
2677617725797602578879882sp: DPPC_BACFIBacillus firmus OF4 dppC38.873.8286dipeptide transport system
permease protein
2678617825807072579769939pir: S47696Escherichia coli K12 nikB33.262.0316nickel transport system permease
protein
26796179258241725807111707
26806180258256425845041941
26816181258461325859261314sp: ARGD_CORGLCorynebacterium glutamicum31.463.5411acetylornithine aminotransferase
ATCC 13032 argD
26826182258618025877631584pir: A70539Mycobacterium tuberculosis25.147.9482hypothetical protein
H37Rv Rv1128c
2683618325879762588722747sp: YA26_MYCTUMycobacterium tuberculosis49.179.4218hypothetical membrane protein
H37Rv Rv0364
2684618425894322588725708sp: PHBB_CHRVIChromatium vinosum D phbB28.160.0235acetoacetyl CoA reductase
2685618525895652590302738pir: A40046Streptomyces coelicolor actII26.755.0240transcriptional regulator, TetR family
2686618625906972591137441GSP: Y74375Neisseria meningitidis38.047.094polypeptides predicted to be useful
antigens for vaccines and
diagnostics
2687618725923652591574792gp: AF106002_1Pseudomonas putida GM7331.165.1238ABC transporter ATP-binding protein
ttg2A
2688618825924022592794393gp: MLCB1610_9Mycobacterium leprae53.277.0126globin
MLCB1610.14c
26896189259283825939651128sp: CHRA_PSEAEPseudomonas aeruginosa27.360.4396chromate transport protein
Plasmid pUM505 chrA
2690619025945942593968627pir: A70867Mycobacterium tuberculosis37.868.9196hypothetical protein
H37Rv Rv2474c
2691619125950612594597465gp: SC6D10_19Streptomyces coelicolor A3(2)36.261.4127hypothetical protein
SC6D10.19c
2692619225958082595188621
2693619325959832595822162pir: B72589Aeropyrum pernix K1 APE118236.460.055hypothetical protein
26946194259771525960481668sp: YJJK_ECOLIEscherichia coli K12 yjjK52.879.6563ABC transporter ATP-binding protein
2695619525984832597869615pir: E70867Mycobacterium tuberculosis31.462.2172hypothetical protein
H37Rv Rv2478c
26966196260076425986622103sp: Y05L_MYCLEMycobacterium leprae o65928.056.7700hypothetical membrane protein
26976197260146126028791419pir: C69676Bacillus subtilis phoB28.052.6536alkaline phosphatase
2698619826045732605502930
2699619926045832603945639
2700620026055202604609912sp: MSMG_STRMUStreptococcus mutans39.176.3279multiple sugar-binding transport
INGBRITT msmGsystem permease protein
2701620126063692605527843sp: MSMF_STRMUStreptococcus mutans27.467.5292multiple sugar-binding transport
INGBRITT msmFsystem permease protein
27026202260644426081171674
27036203260788926065611329prf: 2206392CThermoanaerobacterium28.863.2462maltose-binding protein
thermosul amyE
27046204260942626081851242
27056205261063926095121128prf: 2308356AStreptomyces reticuli msiK59.179.8386ABC transporter ATP-binding protein
(ABC-type sugar transport protein)
or cellobiose/maltose transport
protein
2706620626115232612272750
2707620726115312610848684prf: 2317468ASchizosaccharomyces pombe37.772.7154dolichol phosphate mannose
dpm1synthase
2708620826124622613151690
2709620926137122614500789prf: 2516398ERhodococcus rhodochrous67.289.4207aldehyde dehydrogenase
plasmid pRTL1 orf5
2710621026146492615410762prf: 2513418ASynechococcus sp. PCC794248.673.8183circadian phase modifier
cpmA
2711621126154512615795345
27126212261712026159391182pir: A72312Thermotoga maritima MSB835.064.6412hypothetical membrane protein
TM0964
2713621326172462617995750sp: GIP_ECOLIEscherichia coli K12 gip41.269.4255glyoxylate-induced protein
2714621426180722618869798pir: E70761Mycobacterium tuberculosis40.057.0258ketoacyl reductase
H37Rv Rv1544
2715621526188822619538657sp: ORN_ECOLIEscherichia coli K12 orn48.078.8179oligoribonuclease
27166216262072826195411188prf: 2409378ASalmonella enterica iroD26.050.9454ferric enterochelin esterase
27176217262218126209731209pir: C70870Mycobacterium tuberculosis48.571.9398lipoprotein
H37Rv Rv2518c lppS
2718621826229612623605645
2719621926237702623621150
2720622026238032624048246
27216221262535826240511308gp: SCU53587_1Corynebacterium glutamicum99.599.8436transposase (IS1207)
ATCC 21086
2722622226256002625806207
2723622326264472625809639
2724622426279242628376453gp: AF085239_1Salmonella typhimurium KP100132.863.4131transcriptional regulator
cytR
27256225262812126264931629sp: GLSK_RATRattus norvegicus SPRAGUE-35.269.3358glutaminase
DAWLEY KIDNEY
2726622626283762628852477pir: A36940Bacillus subtilis 168 degA42.372.297sporulation-specific degradation
regulator protein
2727622726288782628324555
27286228262892626304791554sp: UXAC_ECOLIEscherichia coli K12 uxaC29.060.9335uronate isomerase
2729622926306362631136501
27306230263127026324661197prf: 1814452CZea diploperennis perennial32.045.0291hypothetical protein
teosinte
2731623126325432633100558prf: 2324444AMycobacterium avium pncA48.174.6185pyrazinamidase/nicotinamidase
2732623226334182633146273pir: E70870Mycobacterium tuberculosis42.780.075hypothetical protein
H37Rv Rv2520c
2733623326336002634064465sp: BCP_ECOLIEscherichia coli K12 bcp46.873.8141bacterioferritin comigratory protein
2734623426341162634751636gp: SCI11_1Streptomyces coelicolor A3(2)32.561.4114bacterial regulatory protein, tetR
SCI11.01cfamily
2735623526351512634747405gp: BAY15081_1Corynebacterium56.675.9145phosphopantethiene protein
ammoniagenes ATCC 6871 ppt1transferase
27366236263658926351651425gp: AF237667_1Corynebacterium glutamicum52.485.6473lincomycin resistance protein
lmrB
2737623726368452637168324pir: S76537Synechocystis sp. PCC680330.154.0113hypothetical membrane protein
2738623826376532637240414
27396239264762726386498979pir: S2047Corynebacterium62.383.63029fatty-acid synthase
ammoniagenes fas
27406240264941626482351182gp: SC4A7_14Streptomyces coelicolor A3(2)25.355.2404hypothetical protein
SC4A7.14
2741624126495502650164615pir: D70716Mycobacterium tuberculosis40.460.9230peptidase
H37Rv Rv0950c
2742624226504412650902462sp: Y077_MYCTMycobacterium tuberculosis40.267.9112hypothetical membrane protein
H37Rv Rv1343c
2743624326509862651339354sp: Y076_MYCLEMycobacterium leprae37.269.0113hypothetical membrane protein
B1549_F2_59
2744624426520372651420618sp: Y03Q_MYCTUMycobacterium tuberculosis55.076.7202hypothetical protein
H37Rv Rv1341
2745624526528012652067735sp: RNPH_PSEAEPseudomonas aeruginosa60.281.4236ribonuclease PH
ATCC 15692 rph
2746624626532542653009246
2747624726540182653326693
2748624826546602654079582
27496249265623626548751362sp: Y029_MYCTUMycobacterium tuberculosis29.058.2428hypothetical membrane protein
H37Rv SC8A6.09c
2750625026564522656985534gp: AF121000_8Corynebacterium glutamicum92.197.2175transposase (IS1628)
22243 R-plasmid pAG1 tnpB
2751625126576332656974660
2752625226585002657736765sp: Y03O_MYCLEMycobacterium leprae ats46.074.4250arylsulfatase
2753625326594572658606852prf: 2516259ACorynebacterium glutamicum99.399.3284D-glutamate racemase
ATCC 13869 murI
2754625426594962660131636
2755625526606382660147492gp: SCE22_22Streptomyces coelicolor A3(2)44.270.8147bacterial regulatory protein, marR
SCE22.22family
2756625626614172660671747sp: Y03M_MYCTUMycobacterium tuberculosis38.269.3225hypothetical membrane protein
H37Rv Rv1337
2757625726615652662455891
2758625826623762661417960pir: A47039Flavobacterium sp. nylC30.258.3321endo-type 6-aminohexanoate
oligomer hydrolase
2759625926628672662331537sp: Y03H_MYCTUMycobacterium tuberculosis35.058.5200hypothetical protein
H37Rv Rv1332
2760626026631822662883300sp: Y03G_MYCTUMycobacterium tuberculosis57.177.1105hypothetical protein
H37Rv Rv1331
2761626126634372664060624
27626262266406026653971338sp: Y03F_MYCTUMycobacterium tuberculosis61.280.8428hypothetical protein
H37Rv Rv1330c
2763626326656872665992306
27646264266611526678541740prf: 1816252AEscherichia coli dinG25.253.3647ATP-dependent helicase
2765626526687602667870891sp: Y0A8_MYCTUMycobacterium tuberculosis29.760.1313hypothetical membrane protein
H37Rv Rv2560
2766626626695612668839723pir: T34684Streptomyces coelicolor A3(2)39.052.0222hypothetical protein
SC1B5.06c
27676267267057326695571017sp: SERB_ECOLIEscherichia coli K12 serB38.761.0310phosphoserine phosphatase
27686268267112626727211596
27696269267280526710631743pir: D45335Mycobacterium tuberculosis46.874.4575cytochrome c oxidase chain I
H37Rv Rv3043c
2770627026729502673255306
27716271267433926733381002gp: AF112536_1Corynebacterium glutamicum99.799.7334ribonucleotide reductase beta-chain
ATCC 13032 nrdF
2772627226748042675289486sp: FTNA_ECOLIEscherichia coli K12 ftnA31.564.2159ferritin
2773627326754912676240750gp: SCA32WHIH_4Streptomyces coelicolor A3(2)32.860.2256sporulation transcription factor
whiH
2774627426769022676243660pir: I40339Corynebacterium glutamicum27.660.4225iron dependent repressor or
ATCC 13869 dtxRdiptheria toxin repressor
2775627526769402677377438sp: TIR2_YEASTSaccharomyces cerevisiae24.262.1124cold shock protein TIR2 precursor
YPH148 YOR010C TIR2
2776627626771932676918276pir: C69281Archaeoglobus fulgidus AF025150.086.050hypothetical membrane protein
27776277267959826774782121gp: AF112535_3Corynebacterium glutamicum99.9100.0707ribonucleotide reductase alpha-
ATCC 13032 nrdEchain
2778627826804702680784315
2779627926813632681223141SP: RL36_RICPRRickettsia prowazekii58.079.04150S ribosomal protein L36
2780628026815462682376831sp: NADE_BACSUBacillus subtilis 168 nadE55.678.1279NH3-dependent NAD(+) synthetase
278162812681556268146493
2782628226831192683616498
2783628326831252682379747pir: S76790Synechocystis sp. PCC680330.756.4257hypothetical protein
str1563
2784628426834182683131288pir: G70922Mycobacterium tuberculosis41.768.896hypothetical protein
H37Rv Rv3129
27856285268464626836271020sp: ADH2_BACSTBacillus stearothermophilus26.152.8337alcohol dehydrogenase
DSM 2334 adh
27866286268491926862891371sp: MMGE_BACSUBacillus subtilis 168 mmgE27.056.0459Bacillus subtilis mmg (for mother cell
metabolic genes)
2787628726863152687148834pir: T05174Arabidopsis thaliana T6K22.5033.866.2284hypothetical protein
2788628826882402687449792
27896289269005026883891662sp: PGMU_ECOLIEscherichia coli K12 pgm61.780.6556phosphoglucomutase
2790629026901502690437288pir: F70650Mycobacterium tuberculosis41.764.384hypothetical membrane protein
H37Rv Rv3069
2791629126904372690760324pir: D71843Helicobacter pylori J99 jhp114625.461.5122hypothetical membrane protein
2792629226907732691564792sp: YCSI_BACSUBacillus subtilis 168 ycsI51.279.1254hypothetical protein
27936293269168926930531365gp: AF126281_1Rhodococcus erythropolis 24.248.6496transposase (IS1676)
27946294269329926949181620sp: CSP1_CORGLCorynebacterium glutamicum24.849.6355major secreted protein PS1 protein
(Brevibacterium flavum) ATCCprecursor
17965 csp1
2795629526949262695279354
2796629626955542695718165
2797629726957662695320447
27986298269581226972121401gp: AF126281_1Rhodococcus erythropolis 24.646.6500transposase (IS1676)
2799629926981502697383768
28006300269953126981941338sp: GLTT_BACCABacillus subtilis 16830.866.2438proton/sodium-glutamate symport
protein
2801630127009202701612693
28026302270246626999262541gp: SCE25_30Streptomyces coelicolor A3(2)33.069.0873ABC transporter
SCE25.30
2803630327024662703356891
2804630427031942702487708gp: SAU18641_2Staphylococcus aureus 45.479.8218ABC transporter ATP-binding protein
2805630527043142704586273PIR: F81516Chlamydophila pneumoniae60.067.084hypothetical protein
AR39 CP0987
2806630627048352704975141PIR: F81737Chlamydia muridarum Nigg71.075.042hypothetical protein
TC0129
2807630727098782710555678
2808630827106372711308672prf: 2509388LStreptomyces collinus Tu 189228.154.1196oxidoreductase or dehydrogenase
ansG
2809630927118502712374525sp: Y089_MYCTUMycobacterium tuberculosis25.951.2205methyltransferase
H37Rv Rv0089
2810631027131812713453273GSP: Y35814Chlamydia pneumoniae61.066.084hypothetical protein
2811631127137022713842141PIR: F81737Chlamydia muridarum Nigg71.075.042hypothetical protein
TC0129
2812631227181872717993195
28136313271968927184361254sp: MURA_ACICAAcinetobacter calcoaceticus44.875.3417UDP-N-acetylglucosamine 1-
NCIB 8250 murAcarboxyvinyltransferase
2814631427197502720319570sp: Y02Y_MYCTUMycobacterium tuberculosis66.384.2190hypothetical protein
H37Rv Rv1314c
2815631527212272720385843gp: SC2G5_15Streptomyces coelicolor A3(2)45.969.0281transcriptional regulator
SC2G5.15c
2816631627217022721295408
2817631727219342722857924sp: CYSK_BACSUBacillus subtilis 168 cysK57.184.6305cysteine synthase
2818631827230642723609546prf: 2417357CAzotobacter vinelandii cysE261.179.7172O-acetylserine synthase
2819631927240572723770288gp: AE002024_10Deinococcus radiodurans R136.165.183hypothetical protein
DR1844
2820632027253592724478882sp: SUCD_COXBUCoxiella burnetii Nine Mile Ph I52.979.4291succinyl-CoA synthetase alpha
sucDchain
2821632127256192725843225PIR: F72706Aeropyrum pernix K1 APE106942.043.075hypothetical protein
28226322272657727253841194sp: SUCC_BACSUBacillus subtilis 168 sucC39.873.0400succinyl-CoA synthetase beta chain
2823632327271452726786360
2824632427281332727399735gp: AF058302_5Streptomyces roseofulvus frnE38.571.8213frenolicin gene E product
2825632527290252728207819
28266326273091627293781539sp: CAT1_CLOKLClostridium kluyveri cat1 cat147.977.8501succinyl-CoA coenzyme A
transferase
28276327273137627325181143sp: NIR3_AZOBRAzospirillum brasilense ATCC38.668.5321transcriptional regulator
29145 ntrC
2828632827322302731424807
2829632927326362733367732pir: E70810Mycobacterium tuberculosis46.581.7213phosphate transport system
H37Rv Rv0821c phoY-2regulatory protein
2830633027343512733455897pir: S68595Pseudomonas aeruginosa pstB58.882.8255phosphate-specific transport
component
2831633127351842734264921gp: MTPSTA1_1Mycobacterium tuberculosis51.482.2292phosphate ABC transport system
H37Rv Rv0830 pstA1permease protein
28326332273621527352021014pir: A70584Mycobacterium tuberculosis50.278.5325phosphate ABC transport system
H37Rv Rv0829 pstC2permease protein
28336333273753827364141125pir: H70583Mycobacterium tuberculosis40.056.0369phosphate-binding protein S-3
H37Rv phoS2precursor
2834633427387112737836876gp: SCD84_18Streptomyces coelicolor A3(2)34.360.0315acetyltransferase
SCD84.18c
2835633527387712739553783
28366336274065027395561095sp: BMRU_BACSUBacillus subtilis 168 bmrU24.755.2344hypothetical protein
2837633727406702741356687pir: E70809Mycobacterium tuberculosis44.974.2225hypothetical protein
H37Rv Rv0813c
2838633827425772741636942gp: AF193846_1Solanum tuberosum BCAT228.656.0259branched-chain amino acid
aminotransferase
28396339274268527437851101gp: AB003158_6Corynebacterium58.579.0352hypothetical protein
ammoniagenes ATCC 6872
ORF4
2840634027440102744222213pir: B70809Mycobacterium tuberculosis58.681.058hypothetical protein
H37Rv Rv0810c
28416341274595427448811074gp: AB003158_5Corynebacterium81.094.23475′-phosphoribosyl-5-aminoimidazole
ammoniagenes ATCC 6872synthetase
purM
28426342274756427460831482gp: AB003158_4Corynebacterium70.389.0482amidophosphoribosyl transferase
ammoniagenes ATCC 6872
purF
2843634327480572747683375pir: H70536Mycobacterium tuberculosis57.375.8124hypothetical protein
H37Rv Rv0807
28446344274809527491111017gp: AB003158_2Corynebacterium75.994.0315hypothetical protein
ammoniagenes ATCC 6872
ORF2
2845634527499022749162741gp: AB003158_1Corynebacterium67.787.1217hypothetical membrane protein
ammoniagenes ATCC 6872
ORF1
2846634627519182752103186GP: SSU18930_214Sulfolobus solfataricus64.071.042hypothetical protein
28476347275231227500272286gp: AB003162_3Corynebacterium77.689.57635′-phosphoribosyl-N-
ammoniagenes ATCC 6872formylglycinamidine synthetase
purL
2848634827524022753121720
2849634927529952752327669gp: AB003162_2Corynebacterium80.393.32235′-phosphoribosyl-N-
ammoniagenes ATCC 6872formylglycinamidine synthetase
purQ
2850635027532372752995243gp: AB003162_1Corynebacterium81.093.779hypothetical protein
ammoniagenes ATCC 6872
purorf
2851635127532982753819522
2852635227538042753328477prf: 2420329ALactococcus lactis gpo46.277.9158gluthatione peroxidase
28536353275399227567392748prf: 2216389AAeromonas hydrophila JMP63628.051.5965extracellular nuclease
nucH
2854635427568512757126276
2855635527578152757129687pir: C70709Mycobacterium tuberculosis37.468.7211hypothetical protein
H37Rv Rv0784
28566356275920027578631338sp: DCTA_SALTYSalmonella typhimurium LT249.081.6414C4-dicarboxylate transporter
dctA
28576357276164927595322118prf: 2408266APseudomonas sp. WO24 dapb141.870.6697dipeptidyl aminopeptidase
2858635827624522761829624
2859635927626752761785891gp: AB003161_3Corynebacterium70.189.12945′-phosphoribosyl-4-N-
ammoniagenes ATCC 6872succinocarboxamide-5-amino
purCimidazole synthetase
28606360276493127635041428gp: AB003161_2Corynebacterium85.395.0477adenylosuccino lyase
ammoniagenes ATCC 6872
purB
28616361276613527649781158sp: AAT_SULSOSulfolobus solfataricus ATCC28.162.3395aspartate aminotransferase
49255
28626362276742027661581263gp: AB003161_1Corynebacterium71.186.44255′-phosphoribosylglycinamide
ammoniagenes ATCC 6872synthetase
purD
2863636327675802767993414sp: YHIT_MYCLEMycobacterium leprae u296a53.780.2136histidine triad (HIT) family protein
2864636427681372767703435
2865636527690952768343753pir: S62195Methanosarcina barkeri orf326.856.4243hypothetical protein
28666366277051127691561356sp: DTPT_LACLALactococcus lactis subsp. lactis30.167.6469di-/tripeptide transpoter
dipT
28676367277071427719821269sp: BIOA_CORGLCorynebacterium glutamicum95.798.8423adenosylmethionine-8-amino-7-
(Brevibacterium flavum) MJ233oxononanoate aminotransferase or
bioA7,8-diaminopelargonic acid
aminotransferase
2868636827719892772660672sp: BIOD_CORGLCorynebacterium glutamicum98.799.6224dethiobiotin synthetase
(Brevibacterium flavum) MJ233
bioD
28696369277409827726441455gp: AF049873_3Lactococcus lactis M71plasmid31.370.5335two-component system sensor
pND306histidine kinase
2870637027748142774110705prf: 2222216AThermotoga maritima drrA42.072.7231two-component system regulatory
protein
2871637127756892774937753sp: TIPA_STRLIStreptomyces lividans tipA37.469.5249transcriptional activator
28726372277687927757401140prf: 2419350AArthrobacter sp. DK-3830.953.9382metal-activated pyridoxal enzyme or
low specificity D-Thr aldolase
28736373277850427767681737gp: ECOPOXB8G_1Escherichia coli K12 poxB46.375.8574pyruvate oxidase
28746374277896527804461482prf: 2212334BStaphylococcus aureus plasmid33.368.9504multidrug efflux protein
pSK23 qacB
2875637527804392780969531sp: YCDC_ECOLIEscherichia coli K12 ycdC30.468.592transcriptional regulator
28766376278099627823151320pir: D70551Mycobacterium tuberculosis45.678.4421hypothetical membrane protein
H37Rv Rv2508c
28776377278448127823402142
2878637827856152784656960gp: AF096929_2Rhodococcus erythropolis SQ134.362.13033-ketosteroid dehydrogenase
kstD1
2879637927863552785651705sp: ALSR_BACSUBacillus subtilis 168 alsR37.169.0232transcriptional regulator, LysR family
2880638027877822788594813pir: C70982Mycobacterium tuberculosis28.452.9278hypothetical protein
H37Rv Rv3298c lpqC
2881638127893992788587813pir: C69862Bacillus subtilis 168 ykrA26.755.6288hypothetical protein
2882638227899352789477459
2883638327901522790550399pir: A45264Oryctolagus cuniculus kidney28.650.7140hypothetical protein
cortex rBAT
28846384279094627924481503pir: B70798Mycobacterium tuberculosis36.064.0464hypothetical membrane protein
H37Rv Rv3737
2885638527925312792857327pir: S41307Streptomyces griseus hrdB32.350.3155transcription initiation factor sigma
28866386279287327943271455sp: TPS1_SCHPOSchizosaccharomyces pombe38.866.7487trehalose-6-phosphate synthase
tps1
2887638727943002794812513
2888638827948702795637768sp: OTSB_ECOLIEscherichia coli K12 otsB27.457.6245trehalose-phosphatase
28896389279674927956761074sp: CCPA_BACMEBacillus megaterium ccpA24.760.2344glucose-resistance amylase
regulator
2890639027968652797806942sp: ZNUA_HAEINHaemophilus influenzae Rd22.446.7353high-affinity zinc uptake system
HI0119 znuAprotein
2891639127978202798509690gp: AF121672_2Staphylococcus aureus 8325-431.463.2223ABC transporter
mreA
2892639227988372799391555pir: E70507Mycobacterium tuberculosis60.087.4135hypothetical membrane protein
H37Rv Rv2060
28936393279953528010341500pir: A69426Archaeoglobus fulgidus 23.452.5303transposase (ISA0963-5)
2894639428011132801313201
28956395280324628015581689gp: AF096929_2Rhodococcus erythropolis SQ132.162.05613-ketosteroid dehydrogenase
kstD1
2896639628039962803250747
2897639728046912804074618pir: B72359Thermotoga maritima MSB834.356.4204lipopolysaccharide biosynthesis
bplAprotein or oxidoreductase or
dehydrogenase
2898639828051102804676435sp: MI2D_BACSUBacillus subtilis 168 idh or iolG35.269.5128dehydrogenase or myo-inositol 2-
dehydrogenase
2899639928059672805113855sp: SHIA_ECOLIEscherichia coli K12 shiA30.567.5292shikimate transport protein
2900640028064412806016426sp: SHIA_ECOLIEscherichia coli K12 shiA43.180.8130shikimate transport protein
2901640128072522806599654gp: SC5A7_19Streptomyces coelicolor A3(2)32.655.7212transcriptional regulator
SC5A7.19c
2902640228083642807426939sp: PT56_YEASTSaccharomyces cerevisiae22.847.3334ribosomal RNA ribose methylase or
YOR201C PET56tRNA/rRNA methyltransferase
29036403280977828083991380sp: SYC_ECOLIEscherichia coli K12 cysS42.268.8464cysteinyl-tRNA synthetase
29046404281180628098241983prf: 2511335CLactococcus lactis sacB47.077.0668PTS system, enzyme II sucrose
protein (sucrose-specific IIABC
component)
29056405281325828119601299gp: AF205034_4Clostridium acetobutylicum35.356.9473sucrose 6-phosphate hydrolase or
ATCC 824 scrBsucrase
2906640628140372813279759sp: NAGB_ECOLIEscherichia coli K12 nagB38.369.4248glucosamine-6-phosphate
isomerase
29076407281523228140811152sp: NAGA_VIBFUVibrio furnissii SR1514 manD30.260.3368N-acetylglucosamine-6-phosphate
deacetylase
2908640828154582816393936sp: DAPA_ECOLIEscherichia coli K12 dapA28.262.1298dihydrodipicolinate synthase
2909640928164092817317909sp: GLK_STRCOStreptomyces coelicolor A3(2)28.757.6321glucokinase
SC6E10.20c glk
2910641028173632818058696prf: 2516292AClostridium perfringens NCTC36.468.6220N-acetylmannosamine-6-phosphate
8798 nanEepimerase
2911641128183132818137177
29126412281956428183501215sp: NANH_MICVIMicromonospora viridifaciens24.850.3439sialidase precursor
ATCC 31146 nadA
2913641328202852819557729gp: AF181498_1Rhizobium etli ansR26.657.2222L-asparagine permease operon
repressor
29146414282058428221911608gp: BFU64514_1Bacillus firmus OF4 dppA22.551.4560dipeptide transporter protein or
heme-binding protein
2915641528223872823337951sp: DPPB_BACFIBacillus firmus OF4 dappB31.964.3342dipeptide transport system
permease protein
29166416282427428253411068sp: OPPD_BACSUBacillus subtilis 168 oppD46.578.3314oligopeptide transport ATP-binding
protein
2917641728253412826156816sp: OPPF_LACLALactococcus lactis oppF43.478.7258oligopeptide transport ATP-binding
protein
2918641828268352826215621sp: RHTB_ECOLIEscherichia coli K12 rhtB28.562.7193homoserine/homoserin lactone
efflux protein or lysE type
translocator
2919641928269222827404483prf: 2309303ABradyrhizobium japonicum lrp31.066.2142leucine-responsive regulatory
protein
2920642028278172827458360
2921642128283832827904480pir: C70607Mycobacterium tuberculosis55.986.2152hypothetical protein
H37Rv Rv3581c
2922642228291462828379768sp: Y18T_MYCTUMycobacterium tuberculosis46.471.5235hypothetical protein
H37Rv Rv3582c
2923642328297492829156594pir: H70803Mycobacterium tuberculosis73.391.1157transcription factor
H37Rv Rv3583c
2924642428300572830779723prf: 2214304AMycobacterium tuberculosis43.570.0223two-component system response
H37Rv Rv3246c mtrAregulator
29256425283077928318941116sp: BAES_ECOLIEscherichia coli K12 baeS29.367.7341two-component system sensor
histidine kinase
2926642628320852832666582
29276427283279028341811392sp: RADA_ECOLIEscherichia coli K12 radA41.574.3463DNA repair protein RadA
29286428283418828352851098sp: YACK_BACSUBacillus subtilis 168 yacK40.373.3345hypothetical protein
2929642928359692835283687pir: D70804Mycobacterium tuberculosis29.453.3231hypothetical protein
H37Rv Rv3587c
29306430283749928360481452gp: PPU96338_1Pseudomonas putida NCIMB59.585.1471p-hydroxybenzaldehyde
9866 plasmid pRA4000dehydrogenase
2931643128377372837591147
2932643228385762837956621pir: T08204Chlamydomonas reinhardtii ca136.766.2210mitochondrial carbonate
dehydratase beta
2933643328386432839521879gp: AF121797_1Streptomyces antibioticus IMRU48.470.7283A/G-specific adenine glycosylase
3720 mutY
29346434283956228407161155
2935643528410632840758306
2936643628410752841848774gp: AB009078_1Brevibacterium saccharolyticum99.299.6258L-2.3-butanediol dehydrogenase
2937643728421302842453324
2938643828424932843233741
2939643928434052843716312
2940644028437222843432291pir: E70552Mycobacterium tuberculosis48.569.197hypothetical protein
H37Rv Rv3592
2941644128451392845558420GSP: Y29188Pseudomonas aeruginosa57.063.099virulence factor
ORF24222
2942644228458892846101213GSP: Y29193Pseudomonas aeruginosa54.055.072virulence factor
ORF25110
2943644328461862846506321GSP: Y29193Pseudomonas aeruginosa74.075.055virulence factor
ORF25110
29446444284694028441662775sp: MECB_BACSUBacillus subtilis 168 mecB58.586.2832ClpC adenosine triphosphatase/
ATP-binding proteinase
29456445284722928486591431gp: AB035643_1Bacillus cereus ts-4 impdh37.170.2469inosine monophosphate
dehydrogenase
29466446284876928497791011pir: JC6117Rhodococcus rhodochrous nitR24.762.7316transcription factor
29476447285003128518151785sp: PH2M_TRICUTrichosporon cutaneum ATCC33.560.9680phenol 2-monooxygenase
46490
29486448285201728537321716
29496449285376928557091941
29506450285579528575161722
2951645128590442859205162
29526452285905528576131443gp: AF237667_1Corynebacterium glutamicum100.0100.0481lincomycin resistance protein
lmrB
2953645328601452859195951pir: G70807Mycobacterium tuberculosis26.755.8240hypothetical protein
H37Rv Rv3517
29546454286208228605051578gp: AB012100_1Bacillus stearothermophilus lysS41.771.2511lysyl-tRNA synthetase
2955645528629292862132798gp: CGPAN_2Corynebacterium glutamicum29.952.6268pantoate—beta-alanine ligase
ATCC 13032 panC
2956645628636212862929693
2957645728644212863624798
2958645828648482864384465gp: MLCB2548_4Mycobacterium leprae29.069.6138hypothetical membrane protein
MLCB2548.04c
2959645928653432864867477sp: HPPK_METEXMethylobacterium extorquens42.469.01582-amino-4-hydroxy-6-
AM1 folKhydroxymethyldihydropteridine
pyrophosphokinase
2960646028657352865346390sp: FOLB_BACSUBacillus subtilis 168 folB38.169.5118dihydroneopterin aldolase
2961646128665672865731837gp: AB028656_1Mycobacterium leprae folP51.575.0268dihydropteroate synthase
2962646228671732866586588sp: GCH1_BACSUBacillus subtilis 168 mtrA60.686.2188GTP cyclohydrolase I
2963646328674712868385915
2964646428697482867169258056.069.0782cell division protein FtsH
2965646528704442869863582gp: AF008931_1Salmonella typhimurium GP66051.583.0165hypoxanthine
hprtphosphoribosyltransferase
2966646628713892870499891sp: YZC5_MYCTUMycobacterium tuberculosis41.066.8310cell cycle protein MesJ or cytosine
H37Rv Rv3625cdeaminase-related protein
29676467287267728714451233sp: DAC_ACTSPActinomadura sp. R39 dac27.251.4459D-alanyl-D-alanine
carboxypeptidase
2968646828729262873399474sp: IPYR_ECOLIEscherichia coli K12 ppa49.773.6159inorganic pyrophosphatase
2969646928736112873393219
29706470287544328739051539pir: H70886Mycobacterium tuberculosis56.080.7507spermidine synthase
H37Rv speE
2971647128758322875434399sp: Y0B1_MYCTUMycobacterium tuberculosis38.686.4132hypothetical membrane protein
H37Rv Rv2600
2972647228762802875870411sp: Y0B2_MYCTUMycobacterium tuberculosis36.863.2144hypothetical protein
H37Rv Rv2599
2973647328767772876280498sp: Y0B3_MYCTUMycobacterium tuberculosis36.460.1173hypothetical protein
H37Rv Rv2598
2974647428773852876777609sp: Y0B4_MYCTUMycobacterium tuberculosis44.672.3202hypothetical protein
H37Rv Rv2597
2975647528777032877455249sp: PTBA_BACSUBacillus subtilis 168 bgIP30.359.689PTS system, beta-glucosides-
permease II ABC component
2976647628778582877595264
29776477287971028784781233gp: AB017795_2Nocardioides sp. KP7 phdD38.069.6411ferredoxin reductase
2978647828799652880252288gp: SCH69_9Streptomyces coelicolor A3(2)46.473.297hypothetical protein
SCH69.09c
2979647928805442880987444prf: 2516298UBurkholderia pseudomallei ORFE26.759.3135bacterial regulatory protein, marR
family
29806480288099828848823885prf: 2413335AStreptomyces roseosporus cpsB28.451.61241peptide synthase
29816481288330428818441461
29826482288649728849351563prf: 2310295AEscherichia coli K12 padA35.063.7488phenylacetaldehyde dehydrogenase
2983648328878332886916918gp: CJ11168X2_254Campylobacter jejuni Cj060457.379.7241hypothetical protein
2984648428901852890346162GP: MSGTCWPA_1Mycobacterium tuberculosis62.063.054hypothetical protein
2985648528903772890553177GP: MSGTCWPA_1Mycobacterium tuberculosis74.080.031hypothetical protein
29866486289054028888971644gsp: R94368Brevibacterium flavum MJ-23399.5100.0548heat shock protein or chaperon or
groEL protein
2987648728909302890751180
29886488289213828909301209
2989648928931002892138963
29906490289508528931001986
29916491289752528950722454
29926492290032628975282799
29936493290392029003303591prf: 2309326AHomo sapiens MUC5B21.742.31236hypothetical protein
29946494290673829039642775
2995649529072502906639612
29966496290751529088851371pir: G70870Mycobacterium tuberculosis37.168.0447peptidase
H37Rv Rv2522c
2997649729092102909788579
2998649829098302909231600
29996499291017229132283057prf: 2504285BStaphylococcus aureus mnhA35.668.3797Na+/H+ antiporter or multiple
resistance and pH regulation related
protein A or NADH dehydrogenase
3000650029132352913723489gp: AF097740_3Bacillus firmus OF4 mrpC44.281.7104Na+/H+ antiporter or multiple
resistance and pH regulation related
protein C or cation transport system
protein
30016501291374929154161668gp: AF097740_4Bacillus firmus OF4 mrpD35.272.1523Na+/H+ antiporter or multiple
resistance and pH regulation related
protein D
3002650229154822915922441gp: AF097740_5Bacillus firmus OF4 mrpE26.760.9161Na+/H+ antiporter or multiple
resistance and pH regulation related
protein E
3003650329159292916201273prf: 2416476GRhizobium meliloti phaF32.566.277K+ efflux system or multiple
resistance and pH regulation related
protein F
3004650429162052916582378prf: 2504285HStaphylococcus aureus mnhG25.663.6121Na+/H+ antiporter or multiple
resistance and pH regulation related
protein G
3005650529176172917024594pir: D70594Mycobacterium tuberculosis24.754.5178hypothetical protein
H37Rv lipV
30066506291875729176301128sp: YBDK_ECOLIEscherichia coli K12 ybdK27.061.7334hypothetical protein
3007650729194812918819663
3008650829197152920293579sp: DEF_BACSUBacillus subtilis 168 def37.560.9184polypeptide deformylase
3009650929197412919490252pir: D70631Mycobacterium tuberculosis47.970.471hypothetical protein
H37Rv Rv0430
30106510292028629212901005pir: B70631Mycobacterium tuberculosis31.354.2339acetyltransferase (GNAT) family or
H37Rv Rv0428cN terminal acetylating enzyme
3011651129204762919808669
3012651229208492920220630
3013651329213202922108789gp: AF108767_1Salmonella typhimurium LT230.859.931exodeoxyrlbonuclease III or
xthAexonuclease
30146514292211829236171500gp: BFU88888_2Bacillus firmus OF4 cls27.962.0513cardiolipin synthase
3015651529241912924844654
30166516292514729239541194sp: BCR_ECOLIEscherichia coli K12 bcr31.667.2393membrane transport protein or
bicyclomycin resistance protein
30176517292554129267041164gp: VCAJ10968_1Vibrio cholerae JS1569 nptA28.568.9382sodium dependent phosphate pump
3018651829275462926707840sp: PHZC_PSEARPseudomonas aureofaciens 30-8438.856.4289phenazine biosynthesis protein
phzC
3019651929282832927651633
3020652029283182927551768gp: SCE8_16Streptomyces coelicolor A3(2)24.360.8255ABC transporter
SCE8.16c
3021652129292372928302936sp: BCRA_BACLIBacillus licheniformis ATCC36.966.3309ABC transporter ATP-binding protein
9945A bcrA
3022652229297562929256501pir: C70629Mycobacterium tuberculosis47.668.5168mutator mutT protein
H37Rv Rv0413
30236523292995129313361386pir: B70629Mycobacterium tuberculosis35.070.2423hypothetical membrane protein
H37Rv Rv0412c
30246524293134029323711032sp: GLNH_BACSTBacillus stearothermophilus31.564.8270glutamine-binding protein precursor
NUB36 glnH
30256525293257729348292253plr: H70628Mycobacterium tuberculosis41.263.5805serine/threonine kinase
H37Rv Rv0410c pknG
3026652629333982932652747
30276527293840329397671365sp: ADRO_BOVINBos taurus37.267.8457ferredoxin/ferredoxin-NADP
reductase
3028652829399072940452546sp: ELAA_ECOLIEscherichia coli K12 elaA34.060.3156acetyltransferase (GNAT) family
30296529294150829404471062
30306530294250029414721029
3031653129430072942609399
30326532294420529430121194sp: PURT_BACSUBacillus subtills 168 purT59.182.6379phosphoribosylglycinamide
formyltransferase
3033653329465262945639888
3034653429475912946698894pir: S60890Corynebacterium glutamicum77.690.9295insertion element (IS3 related)
orf2
3035653529478862947620267pir: S60889Corynebacterium glutamicum67.484.389insertion element (IS3 related)
orf1
30366536294918829480491140gp: AB016841_1Streptomyces thermoviolaceus22.451.3349two-component system sensor
opc-520 chiShistidine kinase
3037653729498822949265618sp: DEGU_BACBRBacillus brevis ALK36 degU31.765.6218transcriptional regulator
3038653829502072950431225
30396539295172329504341290gp: AB003160_1Corynebacterium89.795.3427adenylosuccinate synthetase
ammoniagenes purA
3040654029519332952691759pir: G70575Mycobacterium tuberculosis34.359.3204hypothetical protein
H37Rv Rv0358
3041654129527092952972264
30426542295414129529751167sp: YFDA_CORGLCorynebacterium glutamicum100.0100.0359hypothetical membrane protein
AS019 ATCC 13059 ORF3
30436543295527229542411032pir: S09283Corynebacterium glutamicum99.7100.0344fructose-bisphosphate aldolase
AS019 ATCC 13059 fda
3044654429564732955523951gp: CGFDA_1Corynebacterium glutamicum100.0100.0304hypothetical protein
AS019 ATCC 13059 ORF1
3045654529574472956830618pir: G70833Mycobacterium tuberculosis76.991.2182methyltransferase
H37Rv Rv0380c
3046654629580362957485552gp: AF058713_1Pyrococcus abyssi pyrE39.165.5174orotate phosphoribosyltransferase
3047654729591102958139972pir: B70834Mycobacterium tuberculosis27.660.0250hypothetical protein
H37Rv Rv0383c
3048654829603712959520852sp: THTM_HUMANHomo sapiens mpsT29.656.12943-mercaptopyruvate
sulfurtransferase
3049654929611872960468720
3050655029630082962730279
3051655129635962963198399
3052655229642582964434177GSP: Y29188Pseudomonas aeruginosa76.082.059virulence factor
ORF24222
3053655329650762965837762GSP: Y29182Pseudomonas aeruginosa38.055.0200virulence factor
ORF23228
3054655429651882965583396GSP: Y29193Pseudomonas aeruginosa62.063.0132virulence factor
ORF25110
30556555296780429664581347pir: S76683Synechocystis sp. PCC680324.754.8489sodium/glutamate symport carrier
slr0625protein
3056655629684032968789387sp: CADF_STAAUStaphylococcus aureus cadC37.071.3108cadmium resistance protein
3057655729689512969808858pir: H75109Pyrococcus abyssi Orsay23.763.3283cation efflux system protein
PAB0462(zinc/cadmium)
30586558296983429710031170gp: AB010439_1Rhodococcus rhodochrous22.545.4476monooxygenase or oxidoreductase
IFO3338or steroid monooxygenase
30596559297101729720571041sp: LUXA_KRYASKryptophanaron alfredi symbiont21.147.4399alkanal monooxygenase alpha chain
luxA
3060656029720992971338762
30616561297320529720601146sp: METB_ECOLIEscherichia coli K12 metB36.562.4375cystathionine gamma-lyase
3062656229737962973230567gp: SC1A2_11Streptomyces coelicolor A3(2)40.267.9184bacterial regulatory protein, lacl
SC1A2.11family
3063656329739612974200240gp: SCE20_34Streptomyces coelicolor A3(2)49.465.289rifampin ADP-ribosyl transferase
SCE20.34c arr
3064656429742002974382183gp: SCE20_34Streptomyces coelicolor A3(2)73.287.556rifampin ADP-ribosyl transferase
SCE20.34c arr
30656565297446729755911125pir: E70812Mycobacterium tuberculosis30.556.2361hypothetical protein
H37Rv Rv0837c
3066656629756292976360732pir: D70812Mycobacterium tuberculosis33.864.7204hypothetical protein
H37Rv Rv0836c
30676567297659629777741179pir: D70834Mycobacterium tuberculosis31.960.6386oxidoreductase
H37Rv Rv0385
3068656829786442977847798pir: B69109Methanobacterium32.067.3275N-carbamoyl-D-amino acid
thermoautotrophicum Delta Hamidohydrolase
MTH1811
3069656929787372978979243
30706570297898229801151134gp: SC4A7_3Streptomyces coelicolor A3(2)28.055.4289hypothetical protein
SC4A7.03
3071657129808872981216330GP: ABCARRA_2Azospirillum brasilense carR38.044.0108novel two-component regulatory
system
30726572298169829801811518prf: 2104333DRhodococcus erythropolis thcA69.690.3507aldehyde dehydrogenase
3073657329824602982023438gp: SAU43299_2Streptomyces albus G hspR47.470.4135heat shock transcription regulator
30746574298367929824951185sp: DNAJ_MYCTUMycobacterium tuberculosis56.780.1397heat shock protein dnaJ
H37Rv RV0352 dnaJ
3075657529845222983887636sp: GRPE_STRCOStreptomyces coelicolor grpE38.766.5212nucleotide exchange factor grpE
protein bound to the ATPase domain
of the molecular chaperone DnaK
30766576298639729845441854gsp: R94587Brevibacterium flavum MJ-23399.899.8618heat shock protein dnaK
dnaK
30776577298683329881641332gp: SCF6_8Streptomyces coelicolor A3(2)42.679.0338hypothetical membrane protein
SCF6.09
3078657829888462988214633sp: PFS_HELPYHelicobacter pylori HP0089 mtn27.260.01955′-methylthioadenosine
nucleosidase and S-
adenosylhomocysteine nucleosidase
30796579299004529888461200
3080658029917182992602885
30816581299328629899543333sp: CUT3_SCHPOSchizosaccharomyces pombe18.948.41311chromosome segregation protein
cut3
3082658229939212993286636
30836583299540529939211485
30846584299678129957471035sp: ADH2_BACSTBacillus stearothermophilus50.081.7334alcohol dehydrogenase
DSM 2334 adh
3085658529971512997366216
3086658629976872997481207
3087658729976882997876189
3088658829982232997963261
3089658929994542998528927pir: F69997Bacillus subtilis ytnM43.570.1301hypothetical membrane protein
3090659030002002999478723gp: SC7A8_10Streptomyces coelicolor A3(2)32.553.2252hypothetical protein
SC7A8.10c
3091659130015123002426915
30926592300153930002411299sp: CYSN_ECOLIEscherichia coli K12 cysN47.378.3414sulfate adenylyltransferase, subunit 1
3093659330024533001542912sp: CYSD_ECOLIEscherichia coli K12 cysD46.170.1308sulfate adenylyltransferase small
chain
3094659430031453002453693sp: CYH1_BACSUBacillus subtilis cysH39.264.2212phosphoadenosine phosphosulfate
reductase
30956595300516230034801683sp: NIR_SYNP7Synechococcus sp. PCC 794234.565.5502ferredoxin—nitrate reductase
30966596300554530069151371sp: ADRO_YEASTSaccharomyces cerevisiae30.861.4487ferredoxin/ferredoxin-NADP
FL200 arh1reductase
30976597300729430083761083prf: 2420294JHomo sapiens hypE32.659.7144huntingtin interactor
3098659830086893008453237
3099659930087703009303534
3100660030091623008749414sp: PHNB_ECOLIEscherichia coli K12 phnB26.859.9142alkylphosphonate uptake protein
and C-P lyase activity
3101660130092423009607366gp: SCE68_10Streptomyces coelicolor A3(2)50.066.380hypothetical protein
SCE68.10
3102660230102313009710522gp: PPAMOA_1Pseudomonas putida DSMZ ID39.176.4161ammonia monooxygenase
88-260 amoA
3103660330106593010979321
3104660430109263010441486
3105660530109893011273285SP: YTZ3_AGRVIAgrobacterium vitis ORFZ341.058.068hypothetical protein
3106660630118053011242564
31076607301280930118081002sp: YGB7_ALCEUAlcaligenes eutrophus H1626.157.9337hypothetical protein
ORF7
3108660830137983013106693gp: HIU68399_3Haemophilus influenzae hmcB35.764.8199ABC transporter
3109660930145503013837714gp: HIU68399_3Haemophilus influenzae hmcB39.373.0211ABC transporter
31106610301461630158241209pir: A69778Bacillus subtilis ydeG30.867.8416metabolite transport protein homolog
3111661130154693014648822
3112661230162383016924687
31136613301714930158271323sp: DAPE_ECOLIEscherichia coli K12 msgB21.548.5466succinyl-diaminopimelate
desuccinylase
31146614301731630192201905
3115661530175393018312774
3116661630181813017420762
3117661730190763018123954GPU: DCA297422_1Daucus carota33.046.0114dehydrin-like protein
31186618302060930195421068sp: MALK_ECOLIEscherichia coli K12 malK24.950.1373maltose/maltodextrin transport ATP-
binding protein
3119661930212023020561642
3120662030218253021208618gp: AF036485_6Lactococcus lactis Plasmid30.267.6179cobalt transport protein
pNZ4000 Orf-200 cbiM
3121662130229283022113816sp: FRP_VIBHAVibrio harveyi MAV frp37.271.4231NADPH-flavin oxidoreductase
3122662230239003022998903sp: IUNH_CRIFACrithidia fasciculata iunH28.459.3317inosine-uridine preferring nucleoside
hydrolase
3123662330243793025353975gp: SCE20_8Streptomyces coelicolor A3(2)31.259.4276hypothetical membrane protein
SCE20.08c
3124662430255523026139588sp: 3MG1_ECOLIEscherichia coli K12 tag50.378.8179DNA-3-methyladenine glycosylase
31256625302729930261421158sp: HMPA_ALCEUAlcaligenes eutrophus H16 fhp33.563.8406flavohemoprotein
3126662630275613028163603
3127662730282683028891624gp: SCO276673_18Streptomyces coelicolor A3(2)34.863.8210oxidoreductase
mmyQ
3128662830288783029033156
3129662930294743028884591sp: BGLG_ECOLIEscherichia coli K12 bglC28.169.3192transcription antiterminator or beta-
glucoside positive regulatory protein
3130663030295043029782279
3131663130300613029702360sp: ABGA_CLOLOClostridium longisporum B640543.759.91676-phospho-beta-glucosidase
abgA
3132663230301553030535381
3133663330303403030101240sp: ABGA_CLOLOClostridium longisporum B640543.978.8666-phospho-beta-glucosidase
abgA
31346634303072330319791257gp: L78665_2Methylobacillus flagellatus aat53.780.9402aspartate aminotransferase
3135663530326473032348300
31366636303266130338631203gp: AF189147_1Corynebacterium glutamicum100.0100.0401transposase (ISCg2)
ATCC 13032 tnp
31376637303418130354371257gp: SCQ11_10Streptomyces coelicolor A3(2)33.670.2399hypothetical membrane protein
SCQ11.10c
3138663830342873034105183
31396639303675630354401317prf: 2422381BSinoRhizobium meliloti rkpK40.572.2442UDP-glucose dehydrogenase
3140664030374113036845567sp: DCD_ECOLIEscherichia coli K12 dcd43.672.3188deoxycytidine triphosphate
deaminase
3141664130376753037911237
3142664230381723038942771gp: SCC75A_16Streptomyces coelicolor A3(2)30.659.4229hypothetical protein
SCC75A.16c
31436643304068130389931689
31446644304193230407481185gp: AB008771_1Streptomyces thermoviolaceus28.558.1410beta-N-Acetylglucosaminidase
nagA
3145664530419943042437444
3146664630425033042703201
31476647304266030457883129gp: MLCB1883_7Mycobacterium leprae29.649.41416hypothetical protein
MLCB1883.13c
3148664830436423043022621
3149664930457963045990195
3150665030471463048048903gp: MLCB1883_4Mycobacterium leprae24.847.1363hypothetical membrane protein
MLCB1883.05c
31516651304718930461221068pir: JC4001Streptomyces sp. acyA27.751.0408acyltransferase or macrolide 3-O-
acyltransferase
3152665230479043047197708
31536653304805830494791422gp: MLCB1883_3Mycobacterium leprae31.254.8529hypothetical membrane protein
MLCB1883.04c
3154665430505223051190669
31556655305059230494561137pir: G70961Mycobacterium tuberculosis53.479.1369hexosyltransferase
H37Rv Rv0225
3156665630511943051964771pir: F70961Mycobacterium tuberculosis58.673.3251methyl transferase
H37Rv Rv0224c
31576657305389130520621830sp: PPCK_NEOFRNeocallimastix frontalis pepck54.778.5601phosphoenolpyruvate carboxykinase
(GTP)
31586658305475930557691011pir: E75125Pyrococcus abyssi Orsay24.452.7332C4-dicarboxylate transporter
PAB2393
3159665930558673056631765sp: YGGH_ECOLIEscherichia coli K12 yggH35.767.2241hypothetical protein
3160666030566133057317705pir: E70959Mycobacterium tuberculosis69.185.0207hypothetical protein
H37Rv Rv0207c
31616661305732830596432316pir: C70839Mycobacterium tuberculosis42.372.3768mebrane transport protein
H37Rv Rv0206c mmpL3
31626662305951730580961422
31636663305965130607331083pir: A70839Mycobacterium tuberculosis29.162.9364hypothetical membrane protein
H37Rv Rv0204c
3164666430607333061095363pir: H70633Mycobacterium tuberculosis34.369.4108hypothetical membrane protein
H37Rv Rv0401
31656665306292730613801548gp: AF113605_1Streptomyces coelicolor A3(2)49.776.9523propionyl-CoA carboxylase complex
pccBB subunit
31666666306778030629514830sp: ERY1_SACERStreptomyces erythraeus eryA30.254.21747polyketide synthase
31676667306993030681431788prf: 2310345AMycobacterium bovis BCG33.562.3592acyl-CoA synthase
3168666830711403070214927pir: F70887Mycobacterium tuberculosis39.867.4319hypothetical protein
H37Rv Rv3802c
3169666930716443071147498
31706670307362030716501971sp: CSP1_CORGLCorynebacterium glutamicum98.699.5657major secreted protein PS1 protein
(Brevibacterium flavum) ATCCprecursor
17965 cop1
31716671307404730754471401
3172667230740753073857219
31736673307656230755401023sp: A85C_MYCTUMycobacterium tuberculosis36.362.5331antigen 85-C
ERDMANN RV0129C fbpC
31746674307877230767152058pir: A70888Mycobacterium tuberculosis37.561.2667hypothetical membrane protein
H37Rv Rv3805c
3175667530798483078853996sp: NOEC_AZOCAAzorhizobium caulinodans27.151.5295nodulation protein
ORS571 noeC
3176667630803513079848504pir: C70888Mycobacterium tuberculosis51.275.0168hypothetical protein
H37Rv Rv3807c
31776677308231130803441968pir: D70888Mycobacterium tuberculosis55.674.7656hypothetical protein
H37Rv Rv3808c
31786678308246730839601494
3179667930844113083935477sp: BCRC_BACLIBacillus licheniformis ATCC28.256.5170phosphatidic acid phosphatase
9945A bcrC
3180668030852003084424777
3181668130857273085218510
31826682308574730870481302sp: FMO1_PIGSus scrofa fmo124.450.4377dimethylaniline monooxygenase (N-
oxide-forming)
3183668330876653088276612
31846684308830330871011203sp: GLF_ECOLIEscherichia coli K12 glf43.272.9377UDP-galactopyranose mutase
31856685308861630906642049pir: G70520Mycobacterium tuberculosis29.647.8659hypothetical protein
H37Rv Rv3811 csp
31866686309228630907601527sp: GLPK_PSEAEPseudomonas aeruginosa51.778.8499glycerol kinase
ATCC 15692 glpK
3187668730931753092342834pir: A70521Mycobacterium tuberculosis41.670.3279hypothetical protein
H37Rv Rv3813c
3188668830940503093175876pir: D70521Mycobacterium tuberculosis46.772.0261acyltransferase
H37Rv Rv3816c
31896689309534330940781266gsp: W26465Mycobacterium tuberculosis70.287.6419seryl-tRNA synthetase
H37Rv
3190669030955743096287714sp: FARR_ECOLIEscherichia coli K12 farR27.761.7235transcriptional regulator, GntR family
or fatty acyl-responsive regulator
31916691309631130974231113pir: H70652Mycobacterium tuberculosis32.661.2356hypothetical protein
H37Rv Rv3835
3192669230974233097764342pir: A70653Mycobacterium tuberculosis46.079.7113hypothetical protein
H37Rv Rv3836
319366933097878309778099
3194669430985723097904669gp: AMU73808_1Amycolatopsis methanolica pgm37.262.82182,3-PDG dependent
phosphoglycerate mutase
3195669530988253099454630
31966696309955631006981143prf: 2501285AMycobacterium smegmatis pzaA27.450.9460nicotinamidase or pyrazinamidase
3197669731006983101426729
31986698310173431027681035gp: SC6G4_33Streptomyces coelicolor A3(2)31.657.1380transcriptional regulator
SC6G4.33
3199669931018633101744120
3200670031026303102079552
3201670131028943103763870
3202670231039263104252327pir: B26872Streptomyces lavendulae43.981.3107hypothetical protein
ORF372
32036703310440631057191314sp: AMYH_YEASTSaccharomyces cerevisiae28.755.3432glucan 1,4-alpha-glucosidase
S288C YIR019C sta1
3204670431069703106053918
3205670531077693106951819sp: GLPQ_BACSUBacillus subtilis glpQ29.054.1259glycerophosphoryl diester
phosphodiesterase
32066706310813131095191389sp: GNTP_BACSUBacillus subtilis gntP37.371.9456gluconate permease
3207670731094643108823642
3208670831098453110003159
32096709311208031104641617sp: KPYK_CORGLCorynebacterium glutamicum25.547.7491pyruvate kinase
AS019 pyk
3210671031133903112449942gsp: Y25997Brevibacterium flavum lctA99.799.7314L-lactate dehydrogenase
32116711311361931153941776pir: C70893Mycobacterium tuberculosis33.564.8526hypothetical protein
H37Rv Rv1069c
3212671231154073116042636gp: SC1C2_30Streptomyces coelicolor A3(2)32.158.5224hydrolase or haloacid
SC1C2.30dehalogenase-like hydrolase
3213671331160793116621543gp: AF030288_1Brevibacterium linens ORF139.967.6188efflux protein
tmpA
3214671431166403117332693sp: GLCC_ECOLIEscherichia coli K12 MG165527.657.0221transcription activator or
glcCtranscriptional regulator GntR family
3215671531173363118121786pir: B70885Mycobacterium tuberculosis47.868.6255phosphoesterase
H37Rv Rv2795c
32166716311828431195821299sp: SHIA_ECOLIEscherichia coli K12 shiA37.974.4422shikimate transport protein
32176717311966531208791215prf: 2219306ANeisseria meningitidis lldA40.468.9376L-lactate dehydrogenase or FMN-
dependent dehydrogenase
3218671831209093121313405
3219671931215983121909312sp: RPC_BPPH1Bacillus phage phi-105 ORF145.580.055immunity repressor protein
3220672031221293121992138
3221672131232223123932711
32226722312417231225561617gp: CELY51B11A_1Caenorhabditis elegans29.551.3569phosphatase or reverse
Y51B11A.1transcriptase (RNA-dependent)
3223672331248863124341546
3224672431252983124897402sp: ILL1_ARATHArabidopsis thaliana ill136.963.1122peptidase or IAA-amino acid
hydrolase
3225672531253433125492150
3226672631261453125495651sp: PMSR_ECOLIEscherichia coli B msrA47.669.1210peptide methionine sulfoxide
reductase
3227672731263923126991600pir: I40858Corynebacterium82.392.7164superoxide dismutase (Fe/Mn)
pseudodiphtheriticum sod
3228672831284173127494924sp: GLTC_BACSUBacillus subtilis gltC32.565.8292transcriptional regulator
32296729312860631297391134gp: AF121000_10Corynebacterium glutamicum23.449.0384multidrug resistance transporter
tetA
32306730312978531313951611
3231673131329203133030111
32326732313302831315081521
3233673331331153133747633pir: G70654Mycobacterium tuberculosis33.864.8216hypothetical protein
H37Rv Rv3850
32346734313526831337781491prf: 2508244ABStreptomyces cyanogenus lanJ27.359.3447membrane transport protein
3235673531352973135752456sp: YXAD_BACSUBacillus subtilis 168 yxaD37.265.0137transcriptional regulator
3236673631364913135856636prf: 2518330BCorynebacterium diphtheriae50.975.5212two-component system response
chrAregulator
3237673731369203137558639
3238673831378843138471588
32396739313790331365931311prf: 2518330ACorynebacterium diphtheriae30.264.5408two-component system sensor
chrShistidine kinase
3240674031386303138481150gp: SCH69_22Streptomyces coelicolor A3(2)45.879.248hypothetical protein
SCH69.22c
3241674131394553138634822gp: SCH69_20Streptomyces coelicolor A3(2)30.059.2277hypothetical protein
SCH69.20c
32426742313965131409521302sp: SP3J_BACSUBacillus subtilis spolllJ26.053.6265stage III sporulation protein
3243674331415233140885639pir: C70948Mycobacterium tuberculosis32.360.9192transcriptional repressor
H37Rv Rv3173c
3244674431419693141709261sp: TAG1_ECOLIEscherichia coli K12.MG165534.571.387transglycosylase-associated protein
tag 1
3245674531433563142454903sp: YW12_MYCTUMycobacterium tuberculosis41.269.6296hypothetical protein
H37Rv Rv2005c
3246674631444823143496987sp: YHBW_ECOLIEscherichia coli K12 MG165538.573.9314hypothetical protein
yhbW
3247674731446613145626966sp: YBC5_CHLVIChlorobium vibrioforme ybc528.451.2334RNA pseudouridylate synthase
3248674831465693146841273GSP: Y35814Chlamydia pneumoniae61.066.084hypothetical protein
3249674931470903147230141PIR: F81737Chlamydia muridarum Nigg71.075.042hypothetical protein
TC0129
3250675031515753151369207
3251675131522043151842363sp: GLCC_ECOLIEscherichia coli K12 MG165530.356.0109bacterial regulatory protein, gntR
glcCfamily or glc operon transcriptional
activator
32526752315241331538281416gp: SC4G6_31Streptomyces coelicolor26.048.2488hypothetical protein
SC4G6.31c
3253675331547663153894873sp: 35KD_MYCTUMycobacterium tuberculosis48.378.7267hypothetical protein
H37Rv Rv2744c
3254675431548173154969153
32556755315669731552461452
32566756315737331563061068
3257675731574713157223249
3258675831577873157479309
3259675931581243158834711gp: SCD35_11Streptomyces coelicolor A3(2)32.358.1217methyltransferase
SCD35.11c
3260676031598003159081720sp: NO21_SOYBNsoybean NO2126.155.2241nodulin 21-related protein
3261676131602163160419204
3262676231606883161065378
3263676331608163161001186
3264676431609383160723216sp: TNP5_PSEAEPseudomonas aeruginosa TNP548.292.956transposon tn501 resolvase
3265676531612193161701483
3266676631614073161087321sp: FER_SACERSaccharopolyspora erythraea fer90.398.462ferredoxin precursor
3267676731620143161682333gp: SCD31_14Streptomyces coelicolor A3(2)47.385.555hypothetical protein
3268676831626943162804111GPU: AF164956_8Corynebacterium glutamicum81.084.027transposase
Tnp1673
3269676931627103162871162GPU: AF164956_23Corynebacterium glutamicum84.090.046transposase protein fragment
TnpNC
32706770316285231638891038
3271677131629833162858126sp: G3P_PYRWOPyrococcus woesei gap63.284.238glyceraldehyde-3-phosphate
dehydrogenase (pseudogene)
3272677231637333163074660pir: S77018Synechocystis sp. PCC680332.259.4180lipoprotein
sll0788
32736773316600531637892217pir: H69268Archaeoglobus fulgidus AF015245.873.4717copper/potassium-transporting
ATPase B or cation transporting
ATPase (E1-E2 family)
3274677431664373166267171
3275677531669783167169192
32766776316764631664501197sp: BAES_ECOLIEscherichia coli K12 baeS37.571.4301two-component system sensor
histidine kinase
3277677731677393168566828
3278677831684013167646756sp: PHOP_BACSUBacillus subtilis phoP43.472.1233two-component response regulator
or alkaline phosphatase synthesis
transcriptional regulatory protein
3279677931686693169340672
32806780316941431708921479sp: COPA_PSESMPseudomonas syringae pv.26.747.9630laccase or copper resistance protein
tomato copAprecursor A
3281678131712543171616363sp: TLPA_BRAJABradyrhizobium japonicum tlpA31.763.4101thiol: disulfide interchange protein
(cytochrome c biogenesis protein)
3282678231725363171619918sp: QOR_MOUSEMus musculus qor31.460.9322quinone oxidoreductase
(NADPH: quinone reductase)(seta-
crystallin)
3283678331729953173465471
3284678431736243173857234sp: ATZN_SYNY3Synechocystis sp. PCC680337.266.778zinc-transporting ATPase (Zn(II)-
atzNtranslocating p-type ATPase
3285678531740663174380315
3286678631749903174784207
32876787317502731769011875sp: ATZN_ECOLIEscherichia coli K12 MG165539.868.5606zinc-transporting ATPase (Zn(II)-
atzNtranslocating p-type ATPase
3288678831756433175254390PIR: E72491Aeropyrum pernix K1 APE257245.054.072hypothetical protein
3289678931771743177482309
3290679031773043177089216GPU: AF164956_8Corynebacterium glutamicum58.073.073transposase
Tnp1673
3291679131775653177308258GPU: AF164956_8Corynebacterium glutamicum75.077.070transposase
Tnp1673
3292679231776833177525159gp: AF121000_8Corynebacterium glutamicum92.596.253transposase (IS1628)
22243 R-plasmid pAG1 tnpB
3293679331785583178112447sp: THI2_ECOLIEscherichia coli K12 thi239.074.0100thioredoxin
3294679431786093178872264
32956795317904931803921344sp: PCAK_PSEPUPseudomonas putida pcaK27.160.1421transmembrane transport protein or
4-hydroxybenzoate transporter
3296679631811043180946159
3297679731811263180551576sp: YQJI_ECOLIEscherichia coli K12 yqjI35.162.5208hypothetical protein
32986798318286631813371530sp: DNAB_ECOLIEscherichia coli K12 dnaB37.773.1461replicative DNA helicase
3299679931834693183984516
3300680031839273183478450sp: RL9_ECOLIEscherichia coli K12 RL942.271.415450S ribosomal protein L9
3301680131846613183987675sp: SSB_ECOLIEscherichia coli K12 ssb30.651.5229single-strand DNA binding protein
3302680231849853184701285sp: RS6_ECOLIEscherichia coli K12 RS628.378.39230S ribosomal protein S6
3303680331855363185348189
33046804318699331855361458gp: AF187306_1Mycobacterium smegmatis41.568.3480hypothetical protein
mc(2)155
3305680531879123188793882
33066806318920131870422160sp: PBPA_BACSUBacillus subtilis ponA29.160.1647penicillin-binding protein
3307680731896523189296357sp: Y0HC_MYCTUMycobacterium tuberculosis41.172.0107hypothetical protein
H37Rv Rv0049
3308680831898773190347471pir: B70912Mycobacterium tuberculosis35.165.0137bacterial regulatory protein, marR
H37Rv Rv0042cfamily
3309680931903783191319942sp: Y0FF_MYCTUMycobacterium tuberculosis29.761.8296hypothetical protein
H37Rv Rv2319c yofF
3310681031913543191848495
3311681131922423191922321sp: YHGC_BACSUBacillus subtilis yhgC32.470.471hypothetical protein
3312681231932013192266936sp: YCEA_ECOLIEscherichia coli K12 yceA30.263.8298hypothetical protein
33136813319451431932521263sp: YBJZ_ECOLIEscherichia coli K12 ybjZ31.264.0433ABC transporter ATP-binding protein
3314681431952033194514690sp: YBJZ_ECOLIEscherichia coli K12 MG165548.980.1221ABC transporter ATP-binding protein
ybjZ
33156815319718631952101977pir: E81408Campylobacter jejuni Cj060618.042.0237hypothetical protein
33166816319741231985001089pir: F70912Mycobacterium tuberculosis77.890.0360hypothetical protein
H37Rv Rv0046c
3317681731991873198582606
33186818320068631992021485
3319681932017543201260495sp: DPS_ECOLIEscherichia coli K12 dps37.764.9154DNA protection during starvation
protein
3320682032019003202712813sp: FPG_ECOLIEscherichia coli K12 mutM or28.455.6268formamidopyrimidine-DNA
fpgglycosylase
33216821320295232041001149sp: RTCB_ECOLIEscherichia coli K12 rtcB47.566.6404hypothetical protein
33226822320406732029791089
3323682332041563204728573
3324682432052043204731474sp: MGMT_HUMANHomo sapiens mgmT38.063.3166methylated-DNA—protein-cysteine
S-methyltransferase
33256825320623232052221011sp: QOR_CAVPOCavia porcellus (Guinea pig) qor33.363.6231zinc-binding dehydrogenase or
quinone oxidoreductase
(NADPH: quinone reductase) or
alginate lyase
3326682632066463206756111
33276827320684932080241176sp: YDEA_ECOLIMycobacterium tuberculosis26.466.3398membrane transport protein
H37Rv Rv0191 ydeA
33286828320827932094541176gp: AF234535_1Corynebacterium melassecola99.799.5392malate oxidoreductase [NAD] (malic
(Corynebacterium glutamicum)enzyme)
ATCC 17965 malE
33296829321118632097051482sp: GNTK_BACSUBacillus subtilis gntK24.553.7486gluconokinase or gluconate kinase
3330683032118363211246591sp: VANZ_ENTFCEnterococcus faecium vanZ27.860.4169teicoplanin resistance protein
3331683132124283211904525sp: VANZ_ENTFCEnterococcus faecium vanZ27.0159.0159teicoplanin resistance protein
33326832321258832139311344sp: MERA_STAAUStaphylococcus aureus merA29.965.6448mercury(II) reductase
33336833321516332139341230sp: DADA_ECOLIEscherichia coli K12 dadA27.354.5444D-amino acid dehydrogenase small
subunit
33346834321675932152571503
3335683532172153216886330
3336683632177773217457321
3337683732179933218601609sp: NOX_THETHThermus thermophilus nox25.855.2194NAD(P)H nitroreductase
3338683832187773219700924
33396839322104432224951452
33406840322263332197782856sp: SYL_BACSUBacillus subtilis syl47.768.1943leucyl-tRNA synthetase
3341684132227223223150429sp: YBAN_ECOLIEscherichia coli K1240.440.4104hypothetical membrane protein
3342684232234453223089357sp: VAPI_BACNODichelobacter nodosus vapI55.881.486virulence-associated protein
3343684332246013225374774
3344684432247143223992723gp: SCC54_19Streptomyces coelicolor31.653.8247hypothetical protein
SCC54.19
3345684532255543224718837sp: HPCE_ECOLIEscherichia coli K12 hpcE28.550.3298bifunctional protein
(homoprotocatechuate catabolism
bifunctional
isomerase/decarboxylase) (2-
hydroxyhepta-2,4-diene-1,7-dioate
isomerase and 5-carboxymethyl-2-
oxo-hex-3-ene-1,7dioate
decarboxylase)
33466846322668732255631125gp: AF173167_1Pseudomonas alcaligenes xlnE34.264.3339gentisate 1,2-dioxygenase or 1-
hydroxy-2-naphthoate dioxygenase
3347684732276893226910780sp: KDGR_ERWCHPectobacterium chrysanthemi25.360.7229bacterial regulatory protein, lacl
kdgRfamily or pectin degradation
repressor protein
33486848322772432290791356sp: PCAK_PSEPUPseudomonas putida pcaK27.560.8454transmembrane transport protein or
4-hydroxybenzoate transporter
33496849322911932304441326prf: 1706191APseudomonas putida 28.249.4476salicylate hydroxylase
33506850323230432310541251sp: EAT2_HUMANHomo sapiens eat225.454.4507proton/glutamate symporter or
excitatory amino acid transporter2
3351685132325963233105510pir: JC2326Corynebacterium glutamicum99.499.4170tryptophan-specific permease
AS019 ORF1
33526852323340332349561554sp: TRPE_BRELABrevibacterium lactofermentum99.299.8515anthranilate synthase component I
trpE
3353685332334203233250171
3354685432349563235579624TRPG_BRELABrevibacterium lactofermentum99.0100.0208anthranilate synthase component II
trpG
33556855323560232366451044sp: TRPD_CORGLCorynebacterium glutamicum99.499.4348anthranilate
ATCC 21850 trpDphosphoribosyltransferase
33566856323664132380621422sp: TRPC_BRELABrevibacterium lactofermentum97.398.3474indole-3-glycerol phosphate
trpCsynthase (IGPS) and N-(5′-
phosphoribosyl) anthranilate
isomerase(PRAI)
3357685732372133236518696
33586858323808232393321251sp: TRPB_BRELABrevibacterium lactofermentum97.697.9417tryptophan synthase beta chain
trpB
3359685932393323240171840sp: TRPA_BRELABrevibacterium lactofermentum95.496.5283tryptophan synthase alpha chain
trpA
33606860324185132403131539gp: SCJ21_17Streptomyces coelicolor A3(2)66.686.8521hypothetical membrane protein
SCJ21.17c
3361686132426883241879810sp: PTXA_ECOLIEscherichia coli K12 ptxA30.371.7152PTS system, IIA component or
unknown pentitol
phosphotransferase enzyme II, A
component
3362686232428543243759906sp: NOSF_PSESTPseudomonas stutzeri32.563.6305ABC transporter ATP-binding protein
33636863324375932453421584gp: SCH10_12Streptomyces coelicolor A3(2)25.257.2547ABC transporter
SCH10.12
3364686432453173245766450sp: UCRI_CHLLTChlorobium limicola petC32.563.6305cytchrome b6-F complex iron-sulfur
subunit (Rieske iron-sulfur protein)
33656865324693132458221110sp: NADO_THEBRThermoanaerobacter brockii33.364.3336NADH oxidase or NADH-dependent
nadOflavin oxidoreductase
3366686632472343248205972sp: YFEH_ECOLIEscherichia coli K12 yfeH43.674.7328hypothetical membrane protein
3367686732483923249165774gp: SCI11_36Streptomyces coelicolor A3(2)34.054.6262hypothetical protein
SCI11.36c
3368686832495343249187348pir: A29606Streptomyces coelicolor Plasmid45.179.4102bacterial regulatory protein, arsR
SCP1 mmrfamily or methylenomycin A
resistance protein
33696869324965132507421092sp: NADO_THEBRThermoanaerobacter brockii33.464.3347NADH oxidase or NADH-dependent
nadOflavin oxidoreductase
3370687032507583251405648sp: YMY0_YEASTSaccharomyces cerevisiae31.469.5226hypothetical protein
ymyO
3371687132516183251466153
3372687232519343251743192
3373687332523003252133168
3374687432526363252316321
3375687532527283253480753sp: BUDC_KLETEKlebsiella terrigena budC26.952.9238acetoin(diacetyl) reductase (acetoin
dehydrogenase)
3376687632535603253739180sp: YY34_MYCTUMycobacterium tuberculosis53.584.558hypothetical protein
H37Rv Rv2094c
33776877325518232538241359sp: DTPT_LACLALactococcus lactis subsp. lactis34.571.6469di-/tripeptide transpoter
dtpT
3378687832555493255719171
3379687932562983255744555sp: ACRR_ECOLIEscherichia coli K12 acrR26.150.5188bacterial regulatory protein, tetR
family
3380688032573733256471903sp: CATA_ACICAAcinetobacter calcoaceticus31.762.2246hydroxyquinol 1,2-dioxygenase
catA
33816881325849132574031089sp: TCBF_PSESQPseudomonas sp. P5143.075.5351maleylacetate reductase
33826882326008432585611524sp: XYLE_ECOLIEscherichia coli K12 xylE31.458.3513sugar transporter or D-xylose-proton
symporter (D-xylose transporter)
3383688332611293261989861sp: ICLR_SALTYSalmonella typhimurium iclR25.760.7280bacterial transcriptional regulator or
acetate operon repressor
33846884326214532632211077sp: YDGJ_ECOLIEscherichia coli K12 ydgJ27.255.7357oxidoreductase
3385688532632373264115879gsp: W61761Listeria innocua strain 445025.958.2270diagnostic fragment protein
sequence
33866886326414232651461005sp: MI2D_BACSUSinoRhizobium meliloti idhA26.559.6332myo-inositol 2-dehydrogenase
33876887326518432662661083sp: STRI_STRGRStreptomyces griseus strI34.162.4343dehydrogenase or myo-inositol 2-
dehydrogenase or streptomycin
biosynthesis protein
33886888326706232710934032pir: C70044Bacillus subtilis yvnB33.362.71242phosphoesterase
3389688932685573267913645
3390689032692353268618618
33916891327139232724771086
3392689232752313274488744sp: UNC1_CAEELCaenorhabditis elegans unc128.657.3206stomatin
3393689332765703275602969
33946894328159932766714929gp: MBO18605_3Mycobacterium bovis BCG58.480.21660DEAD box RNA helicase family
RvD1-Rv2024c
3395689532821723281666507prf: 2323363AAMMycobacterium leprae u2266k34.861.0141hypothetical membrane protein
3396689632827423283101360
3397689732829463282347600sp: THID_BACSUBacillus subtilis thiD50.476.8125phosphomethylpyrimidine kinase
3398689832831413283383243pir: F70041Bacillus subtilis yvgY46.370.167mercuric ion-binding protein or
heavy-metal-associated domain
containing protein
3399689932843093283473837prf: 2501295ACorynebacterium glutamicum29.962.3297ectoine/proline uptake protein
proP
3400690032853553284399957sp: FECB_ECOLIEscherichia coli K12 fecB29.460.6279iron(III) dicitrate-binding periplasmic
protein precursor or iron(III) dicitrate
transport system permease protein
34016901328545532865761122sp: MRF1_SCHPOSchizosaccharomyces pombe27.258.0324mitochondrial respiratory function
mrf1protein or zinc-binding
dehydrogenase or NADPH quinone
oxidoreductase
3402690232866223287005384
3403690332872973287079219
3404690432881903287393798sp: THID_BACSUBacillus subtilis thiD46.275.5249phosphomethylpyrimidine kinase
3405690532882653288609345
3406690632886853288885201pir: F70041Bacillus subtilis yvgY41.870.167mercuric ion-binding protein or
heavy-metal-associated domain
containing protein
3407690732893153288971345sp: AZLD_BACSUBacillus subtilis azlD36.365.7102branched-chain amino acid transport
3408690832900213289311711sp: AZLC_BACSUBacillus subtilis azlD32.167.0212branched-chain amino acid transport
3409690932905913290025567sp: YQGE_ECOLIEscherichia coli K12 yqgE23.756.2169hypothetical protein
34106910329194232906231320sp: CCA_ECOLIEscherichia coli K12 cca26.851.8471tRNA nucleotidyltransferase
3411691132925323293497966pir: E70600Mycobacterium tuberculosis43.669.2234mutator mutT protein
H37Rv Rv3908
3412691232928823292610273
34136913329349732960072511pir: F70600Mycobacterium tuberculosis25.854.3858hypothetical membrane protein
H37Rv Rv3909
34146914329615632994043249pir: G70600Mycobacterium tuberculosis35.760.11201hypothetical membrane protein
H37Rv Rv3910
3415691532977063298428723
3416691632996613300263603sp: RPSH_PSEAEPseudomonas aeruginosa algU30.260.9189RNA polymerase sigma-H factor or
sigma-70 factor (ECF subfamily)
3417691733003713301321951sp: TRXB_STRCLStreptomyces clavuligerus trxB60.482.5308thioredoxin reductase
34186918330130333001191185
3419691933013583301729372sp: THI2_CHLREChlamydomonas reinhardtii thi242.076.5119thioredoxin ch2, M-type
34206920330175533029961242sp: CWLB_BACSUBacillus subtilis cwlB51.075.4196N-acetylmuramoyl-L-alanine
amidase
3421692133027653301989777
34226922330343533044751041
3423692333036163302999618pir: D70851Mycobacterium tuberculosis34.458.5212hypothetical protein
H37Rv Rv3916c
34246924330478733036361152sp: YGI2_PSEPUPseudomonas putida ygi237.660.5367hypothetical protein
3425692533056713304835837sp: YGI1_PSEPUMycobacterium tuberculosis65.078.0272partitioning or sporulation protein
H37Rv parB
3426692633065323305864669sp: GIDB_ECOLIEscherichia coli K12 gidB36.064.7153glucose inhibited division protein B
3427692733076323306682951pir: A70852Mycobacterium tuberculosis44.775.4313hypothetical membrane protein
H37Rv Rv3921c
3428692833083693307971399sp: RNPA_BACSUBacillus subtilis rnpA26.859.4123ribonuclease P protein component
3429692933087473308412336gp: MAU19185_1Mycobacterium avium rpmH83.093.64750S ribosomal protein L34
3430693033090283309321294
3431693133090433308822222
34326932147980147573408gp: AF116184_1Corynebacterium glutamicum100.0100.0136L-aspartate-alpha-decarboxylase
panDprecursor
343369332680012661541848sp: LEU1_CORGLCorynebacterium glutamicum100.0100.06162-isopropylmalate synthase
ATCC 13032 leuA
34346934269068268814255sp: YLEU_CORGLCorynebacterium glutamicum100.0100.085hypothetical protein
(Brevibacterium flavum) ATCC
13032 orfX
343569352706602716911032sp: DHAS_CORGLCorynebacterium glutamicum100.0100.0344aspartate-semialdehyde
asddehydrogenase
34366936446075446521447gp: AF124518_1Corynebacterium glutamicum100.0100.01493-dehydroquinase
ASO19 aroD
343769375263765275631188sp: EFTU_CORGLCorynebacterium glutamicum100.0100.0396elongation factor Tu
ATCC 13059 tuf
343869385694525707711320sp: SECY_CORGLCorynebacterium glutamicum100.0100.0440preprotein translocase secY subuit
(Brevibacterium flavum) MJ233
secY
343969396800446778312214sp: IDH_CORGLCorynebacterium glutamicum100.0100.0738isocitrate dehydrogenase
ATCC 13032 icd(oxalosuccinatedecarboxylase)
344069407203527185801773prf: 2223173ACorynebacterium glutamicum100.0100.0591acyl-CoA carboxylase or biotin-
ATCC 13032 accBCbinding protein
344169418778388791481311sp: CISY_CORGLCorynebacterium glutamicum100.0100.0437citrate synthase
ATCC 13032 gltA
34426942879276879629354sp: FKBP_CORGLCorynebacterium glutamicum100.0100.0118putative binding protein or peptidyl-
ATCC 13032 fkbAprolyl cis-trans isomerase
344369439449969467801785sp: BETP_CORGLCorynebacterium glutamicum100.0100.0595glycine betaine transporter
ATCC 13032 betP
34446944103028310290061278sp: YLI2_CORGLCorynebacterium glutamicum100.0100.0426hypothetical membrane protein
ATCC 13032 orf2
34456945103187110303691503sp: LYSI_CORGLCorynebacterium glutamicum100.0100.0501L-lysine permease
ATCC 13032 lysI
34466946115468311532951389sp: AROP_CORGLCorynebacterium glutamicum100.0100.0463aromatic amino acid permease
ATCC 13032 aroP
3447694711556761154729948pir: S52753Corynebacterium glutamicum100.0100.0316hypothetical protein
ATCC 13032 orf3
34486948115573111568371107prf: 2106301ACorynebacterium glutamicum100.0100.0369succinyl diaminopimelate
ATCC 13032 dapE
34496949121960212180311572gp: CGPUTP_1Corynebacterium glutamicum100.0100.0524proline transport system
ATCC 13032 putP
34506950123827412399231650sp: SYR_CORGLCorynebacterium glutamicum100.0100.0550arginyl-tRNA synthetase
AS019 ATCC 13059 argS
34516951123992912412631335sp: DCDA_CORGLCorynebacterium glutamicum100.0100.0445diaminopimelate (DAP)
AS019 ATCC 13059 lysAdecarboxylase (meso-
diaminopimelate decarboxylase)
34526952124250712438411335sp: DHOM_CORGLCorynebacterium glutamicum100.0100.0445homoserine dehydrogenase
AS019 ATCC 13059 hom
3453695312438551244781927sp: KHSE_CORGLCorynebacterium glutamicum100.0100.0309homoserine kinase
AS019 ATCC 13059 thrB
3454695413276171328243627gsp: W37716Corynebacterium glutamicum100.0100.0216ion channel subunit
R127 orf3
3455695513289531328246708sp: LYSE_CORGLCorynebacterium glutamicum100.0100.0236lysine exporter protein
R127 lysE
3456695613290151329884870sp: LYSG_CORGLCorynebacterium glutamicum100.0100.0290lysine export regulator protein
R127 lysG
34576957133813113400081878sp: ILVB_CORGLCorynebacterium glutamicum100.0100.0626acetohydroxy acid synthase, large
ATCC 13032 ilvBsubunit
3458695813400251340540516pir: B48648Corynebacterium glutamicum100.0100.0172acetohydroxy acid synthase, small
ATCC 13032 ilvNsubunit
34596959134072413417371014pir: C48648Corynebacterium glutamicum100.0100.0338acetohydroxy acid isomeroreductase
ATCC 13032 ilvC
34606960135348913545081020sp: LEU3_CORGLCorynebacterium glutamicum100.0100.03403-isopropylmalate dehydrogenase
ATCC 13032 leuB
34616961142321714252652049prf: 2014259ACorynebacterium glutamicum100.0100.0683PTS system, phosphoenolpyruvate
KCTC1445 ptsMsugar phosphotransferase
(mannose and glucose transport)
3462696214664911467372882sp: ARGB_CORGLCorynebacterium glutamicum100.0100.0294acetylglutamate kinase
ATCC 13032 argB
3463696314685651469521957sp: OTCA_CORGLCorynebacterium glutamicum100.0100.0319ornithine carbamoyltransferase
ATCC 13032 argF
3464696414695281470040513gp: AF041436_1Corynebacterium glutamicum100.0100.0171arginine repressor
ASO19 argR
34656965154455415431541401gp: CGL238250_1Corynebacterium glutamicum100.0100.0467NADH dehydrogenase
ATCC 13032 ndh
3466696615867251586465261gp: AF086704_1Corynebacterium glutamicum100.0100.087phosphoribosyl-ATP-
ASO19 hisEpyrophosphohydrolase
34676967167520816741231086gp: CGL007732_4Corynebacterium glutamicum100.0100.0362ornithine-cyclodecarboxylase
ATCC 13032 ocd
34686968167662316752681356gp: CGL007732_3Corynebacterium glutamicum100.0100.0452ammonium uptake protein, high
ATCC 13032 amtaffinity
3469696916772791677049231gp: CGL007732_2Corynebacterium glutamicum100.0100.077protein-export membrane protein
ATCC 13032 secGsecG
34706970168014316773872757prf: 1509267ACorynebacterium glutamicum100.0100.0919phosphoenolpyruvate carboxylase
ATCC 13032 ppc
34716971172089817196691230gp: AF124600_1Corynebacterium glutamicum100.0100.0410chorismate synthase (5-
AS019 aroCenolpyruvylshikimate-3-phosphate
phospholyase)
34726972188049018823851896pir: B55225Corynebacterium glutamicum100.0100.0632restriction endonuclease
ATCC 13032 cglIIR
3473697320208542021846993prf: 2204286DCorynebacterium glutamicum100.0100.0331sigma factor or RNA polymerase
ATCC 13869 sigBtranscription factor
3474697420606202061504885sp: GLUB_CORGLCorynebacterium glutamicum100.0100.0295glutamate-binding protein
ATCC 13032 gluB
34756975206511620639891128sp: RECA_CORGLCorynebacterium glutamicum100.0100.0376recA protein
AS019 recA
3476697620801832079281903sp: DAPA_BRELACorynebacterium glutamicum100.0100.0301dihydrodipicolinate synthase
(Brevibacterium lactofermentum)
ATCC 13869 dapA
3477697720819342081191744sp: DAPB_CORGLCorynebacterium glutamicum100.0100.0248dihydrodipicolinate reductase
(Brevibacterium lactofermentum)
ATCC 13869 dapB
34786978211536321138641500gp: CGA224946_1Corynebacterium glutamicum100.0100.0500L-malate dehydrogenase (acceptor)
R127 mqo
34796979217174121696662076gp: CAJ10319_4Corynebacterium glutamicum100.0100.0692uridilylyltransferase, uridilylyl-
ATCC 13032 glnDremoving enzyme
3480698021720862171751336gp: CAJ10319_3Corynebacterium glutamicum100.0100.0112nitrogen regulatory protein P-II
ATCC 13032 glnB
34816981217346721721541314gp: CAJ10319_2Corynebacterium glutamicum100.0100.0438ammonium transporter
ATCC 13032 amtP
34826982219608221947421341pir: S32227Corynebacterium glutamicum100.0100.0447glutamate dehydrogenase (NADP+)
ATCC 17965 gdhA
34836983220709222056681425sp: KPYK_CORGLCorynebacterium glutamicum100.0100.0475pyruvate kinase
AS019 pyk
3484698423175502316582969gp: AF096280_1Corynebacterium glutamicum100.0100.0323glucokinase
ATCC 13032 glk
34856985234882923502591431prf: 2322244ACorynebacterium glutamicum100.0100.0477glutamine synthetase
ATCC 13032 glnA
34866986235504223536001443sp: THRC_CORGLCorynebacterium glutamicum100.0100.0481threonine synthase
thrC
34876987245017224483281845prf: 2501295BCorynebacterium glutamicum100.0100.0615ectoine/proline/glycine betaine
ATCC 13032 ectPcarrier
34886988247014124679252217pir: I40715Corynebacterium glutamicum100.0100.0739malate synthase
ATCC 13032 aceB
34896989247074024720351296pir: I40713Corynebacterium glutamicum100.0100.0432isocitrate lyase
ATCC 13032 aceA
34906990249777624966701107sp: PROB_CORGLCorynebacterium glutamicum100.0100.0369glutamate 5-kinase
ATCC 17965 proB
34916991259146925903121158gp: AF126953_1Corynebacterium glutamicum100.0100.0386cystathionine gamma-synthase
ASO19 metB
3492699226801272679684444gp: AF112535_2Corynebacterium glutamicum100.0100.0148ribonucleotide reductase
ATCC 13032 nrdI
3493699326806492680419231gp: AF112535_1Corynebacterium glutamicum100.0100.077glutaredoxin
ATCC 13032 nrdH
3494699427877152786756960sp: DDH_CORGLCorynebacterium glutamicum100.0100.0320meso-diaminopimelate D-
KY10755 ddhdehydrogenase
3495699528880782887944135gp: CGL238703_1Corynebacterium glutamicum100.0100.045porin or cell wall channel forming
MH20-22B porAprotein
34966996293650529353151191sp: ACKA_CORGLCorynebacterium glutamicum100.0100.0397acetate kinase
ATCC 13032 ackA
3497699729374942936508987prf: 2516394ACorynebacterium glutamicum100.0100.0329phosphate acetyltransferase
ATCC 13032 pta
34986998296134229627181377prf: 2309322ACorynebacterium glutamicum100.0100.0459multidrug resistance protein or
ATCC 13032 cmrmacrolide-efflux pump or
drug: proton antiporter
34996999296616129636062556sp: CLPB_CORGLCorynebacterium glutamicum100.0100.0852ATP-dependent protease regulatory
ATCC 13032 clpBsubunit
3500700030995223098578945prf: 1210266ACorynebacterium glutamicum100.0100.0315prephenate dehydratase
pheA
35017001327407432725631512prf: 2501295ACorynebacterium glutamicum100.0100.0504ectoine/proline uptake protein
ATCC 13032 proP
|
EXAMPLE 2
Determination of Effective Mutation Site
(1) Identification of Mutation Site Based on the Comparison of the Gene Nucleotide Sequence of Lysine-Producing B-6 Strain with that of Wild Type Strain ATCC 13032
Corynebacterium glutamicum B-6, which is resistant to S-(2-aminoethyl)cysteine (AEC), rifampicin, streptomycin and 6-azauracil, is a lysine-producing mutant having been mutated and bred by subjecting the wild type ATCC 13032 strain to multiple rounds of random mutagenesis with a mutagen, N-methyl-N′-nitro-N-nitrosoguanidine (NTG) and screening (Appl. Microbiol. Biotechnol., 32: 269-273 (1989)). First, the nucleotide sequences of genes derived from the B-6 strain and considered to relate to the lysine production were determined by a method similar to the above. The genes relating to the lysine production include lysE and lysG which are lysine-excreting genes; ddh, dapA, hom and lysC (encoding diaminopimelate dehydrogenase, dihydropicolinate synthase, homoserine dehydrogenase and aspartokinase, respectively) which are lysine-biosynthetic genes; and pyc and zwf (encoding pyruvate carboxylase and glucose-6-phosphate dehydrogenase, respectively) which are glucose-metabolizing genes. The nucleotide sequences of the genes derived from the production strain were compared with the corresponding nucleotide sequences of the ATCC 13032 strain genome represented by SEQ ID NOS:1 to 3501 and analyzed. As a result, mutation points were observed in many genes. For example, no mutation site was observed in lysE, lysG, ddh, dapA, and the like, whereas amino acid replacement mutations were found in hom, lysC, pyc, zwf, and the like. Among these mutation points, those which are considered to contribute to the production were extracted on the basis of known biochemical or genetic information. Among the mutation points thus extracted, a mutation, Val59Ala, in hom and a mutation, Pro458Ser, in pyc were evaluated whether or not the mutations were effective according to the following method.
(2) Evaluation of Mutation, Val59Ala, in hom and Mutation, Pro458Ser, in pyc
It is known that a mutation in horn inducing requirement or partial requirement for homoserine imparts lysine productivity to a wild type strain (Amino Acid Fermentation, ed. by Hiroshi Aida et al., Japan Scientific Societies Press). However, the relationship between the mutation, Val59Ala, in hom and lysine production is not known. It can be examined whether or not the mutation, Val59Ala, in hom is an effective mutation by introducing the mutation to the wild type strain and examining the lysine productivity of the resulting strain. On the other hand, it can be examined whether or not the mutation, Pro458Ser, in pyc is effective by introducing this mutation into a lysine-producing strain which has a deregulated lysine-bioxynthetic pathway and is free from the pyc mutation, and comparing the lysine productivity of the resulting strain with the parent strain. As such a lysine-producing bacterium, No. 58 strain (FERM BP-7134) was selected (hereinafter referred to the “lysine-producing No. 58 strain” or the “No. 58 strain”). Based on the above, it was determined that the mutation, Val59Ala, in hom and the mutation, Pro458Ser, in pyc were introduced into the wild type strain of Corynebacterium glutamicum ATCC 13032 (hereinafter referred to as the “wild type ATCC 13032 strain” or the “ATCC 13032 strain”) and the lysine-producing No. 58 strain, respectively, using the gene replacement method. A plasmid vector pCES30 for the gene replacement for the introduction was constructed by the following method.
A plasmid vector pCE53 having a kanamycin-resistant gene and being capable of autonomously replicating in Coryneform bacteria (Mol. Gen. Genet., 196: 175-178 (1984)) and a plasmid pMOB3 (ATCC 77282) containing a levansucrase gene (sacB) of Bacillus subtilis (Molecular Microbiology, 6: 1195-1204 (1992)) were each digested with PstI. Then, after agarose gel electrophoresis, a pCE53 fragment and a 2.6 kb DNA fragment containing sacB were each extracted and purified using GENECLEAN Kit (manufactured by BIO 101). The pCE53 fragment and the 2.6 kb DNA fragment were ligated using Ligation Kit ver. 2 (manufactured by Takara Shuzo), introduced into the ATCC 13032 strain by the electroporation method (FEMS Microbiology Letters, 65: 299 (1989)), and cultured on BYG agar medium (medium prepared by adding 10 g of glucose, 20 g of peptone (manufactured by Kyokuto Pharmaceutical), 5 g of yeast extract (manufactured by Difco), and 16 g of Bactoagar (manufactured by Difco) to 1 liter of water, and adjusting its pH to 7.2) containing 25 μg/ml kanamycin at 30° C. for 2 days to obtain a transformant acquiring kanamycin-resistance. As a result of digestion analysis with restriction enzymes, it was confirmed that a plasmid extracted from the resulting transformant by the alkali SDS method had a structure in which the 2.6 kb DNA fragment had been inserted into the PstI site of pCE53. This plasmid was named pCES30.
Next, two genes having a mutation point, hom and pyc, were amplified by PCR, and inserted into pCES30 according to the TA cloning method (Bio Experiment Illustrated vol. 3, published by Shujunsha). Specifically, pCES30 was digested with BamHI (manufactured by Takara Shuzo), subjected to an agarose gel electrophoresis, and extracted and purified using GENECLEAN Kit (manufactured by BIO 101). The both ends of the resulting pCES30 fragment were blunted with DNA Blunting Kit (manufactured by Takara Shuzo) according to the attached protocol. The blunt-ended pCES30 fragment was concentrated by extraction with phenol/chloroform and precipitation with ethanol/and allowed to react in the presence of Taq polymerase (manufactured by Roche Diagnostics) and dTTP at 70° C. for 2 hours so that a nucleotide, thymine (T), was added to the 3′-end to prepare a T vector of pCES30.
Separately, chromosomal DNA was prepared from the lysine-producing B-6 strain according to the method of Saito et al. (Biochem. Biophys. Acta, 72: 619 (1963)). Using the chromosomal DNA as a template, PCR was carried out with Pfu turbo DNA polymelase (manufactured by Stratagene). In the mutated hom gene, the DNAs having the nucleotide sequences represented by SEQ ID NOS:7002 and 7003 were used as the primer set. In the mutated pyc gene, the DNAs having the nucleotide sequences represented by SEQ ID NOS:7004 and 7005 were used as the primer set. The resulting PCR product was subjected to agarose gel electrophoresis, and extracted and purified using GENEGLEAN Kit (manufactured by BIO 101). Then, the PCR product was allowed to react in the presence of Taq polymerase (manufactured by Roche Diagnostics) and dATP at 72° C. for 10 minutes so that a nucleotide, adenine (A), was added to the 3′-end.
The above pCES30 T vector fragment and the mutated hom gene (1.7 kb) or mutated pyc gene (3.6 kb) to which the nucleotide A had been added of the PCR product were concentrated by extraction with phenol/chloroform and precipitation with ethanol, and then ligated using Ligation Kit ver. 2. The ligation products were introduced into the ATCC 13032 strain according to the electroporation method, and cultured on BYG agar medium containing 25 μg/ml kanamycin at 30° C. for 2 days to obtain kanamycin-resistant transformants. Each of the resulting transformants was cultured overnight in BYG liquid medium containing 25 μg/ml kanamycin, and a plasmid was extracted from the culturing solution medium according to the alkali SDS method. As a result of digestion analysis using restriction enzymes, it was confirmed that the plasmid had a structure in which the 1.7 kb or 3.6 kb DNA fragment had been inserted into pCES30. The plasmids thus constructed were named respectively pChom59 and pCpyc458.
The introduction of the mutations to the wild type ATCC 13032 strain and the lysine-producing No. 58 strain according to the gene replacement method was carried out according to the following method. Specifically, pChom59 and pCpyc458 were introduced to the ATCC 13032 strain and the No. 58 strain, respectively, and strains in which the plasmid is integrated into the chromosomal DNA by homologous recombination were selected using the method of Ikeda et al. (Microbiology 144: 1863 (1998)). Then, the stains in which the second homologous recombination was carried out were selected by a selection method, making use of the fact that the Bacillus subtilis levansucrase encoded by pCES30 produced a suicidal substance (J. of Bacteriol., 174: 5462 (1992)). Among the selected strains, strains in which the wild type hom and pyc genes possessed by the ATCC 13032 strain and the No. 58 strain were replaced with the mutated hom and pyc genes, respectively, were isolated. The method is specifically explained below.
One strain was selected from the transformants containing the plasmid, pChom59 or pCpyc458, and the selected strain was cultured in BYG medium containing 20 μg/ml kanamycin, and pCG11 (Japanese Published Examined Patent Application No. 91827/94) was introduced thereinto by the electroporation method. pCG11 is a plasmid vector having a spectinomycin-resistant gene and a replication origin which is the same as pCE53. After introduction of the pCG11, the strain was cultured on BYG agar medium containing 20 μg/ml kanamycin and 100 μg/ml spectinomycin at 30° C. for 2 days to obtain both the kanamycin- and spectinomycin-resistant transformant. The chromosome of one strain of these transformants was examined by the Southern blotting hybridization according to the method reported by Ikeda et al. (Microbiology, 144: 1863 (1998)). As a result, it was confirmed that pChom59 or pCpyc458 had been integrated into the chromosome by the homologous recombination of the Cambell type. In such a strain, the wild type and mutated hom or pyc genes are present closely on the chromosome, and the second homologous recombination is liable to arise therebetween.
Each of these transformants (having been recombined once) was spread on Suc agar medium (medium prepared by adding 100 g of sucrose, 7 g of meat extract, 10 g of peptone, 3 g of sodium chloride, 5 g of yeast extract (manufactured by Difco), and 18 g of Bactoagar (manufactured by Difco) to 1 liter of water, and adjusting its pH 7.2) and cultured at 30° C. for a day. Then the colonies thus growing were selected in each case. Since a strain in which the sacb gene is present converts sucrose into a suicide substrate, it cannot grow in this medium (J. Bacteriol., 174: 5462 (1992)). On the other hand, a strain in which the sacB gene was deleted due to the second homologous recombination between the wild type and the mutated hom or pyc genes positioned closely to each other forms no suicide substrate and, therefore, can grow in this medium. In the homologous recombination, either the wild type gene or the mutated gene is deleted together with the sacB gene. When the wild type is deleted together with the sacb gene, the gene replacement into the mutated type arises.
Chromosomal DNA of each the thus obtained second recombinants was prepared by the above method of Saito et al. PCR was carried out using Pfu turbo DNA polymerase (manufactured by Stratagene) and the attached buffer. In the hom gene, DNAs having the nucleotide sequences represented by SEQ ID NOS:7002 and 7003 were used as the primer set. Also, in the pyc gene was used, DNAs having the nucleotide sequences represented by SEQ ID NOS:7004 and 7005 were used as the primer set. The nucleotide sequences of the PCR products were determined by the conventional method so that it was judged whether the hom or pyc gene of the second recombinant was a wild type or a mutant. As a result, the second recombinant which were called HD-1 and No. 58pyc were target strains having the mutated hom gene and pyc gene, respectively.
(3) Lysine Production Test of HD-1 and No. 58pyc Strains
The HD-1 strain (strain obtained by incorporating the mutation, Val59Ala, in the hom gene into the ATCC 13032 strain) and the No. 58pyc strain (strain obtained by incorporating the mutation, Pro458Ser, in the pyc gene into the lysine-producing No. 58 strain) were subjected to a culture test in a 5 1 jar fermenter by using the ATCC 13032 strain and the lysine-producing No. 58 strain respectively as a control. Thus lysine production was examined.
After culturing on BYG agar medium at 30° C. for 24 hours, each strain was inoculated into 250 ml of a seed medium (medium prepared by adding 50 g of sucrose, 40 g of corn steep liquor, 8.3 g of ammonium sulfate, 1 g of urea, 2 g of potassium dihydrogenphosphate, 0.83 g of magnesium sulfate heptahydrate, 10 mg of iron sulfate heptahydrate, 1 mg of copper sulfate pentahydrate, 10 mg of zinc sulfate hentahydrate, 10 mg of β-alanine, 5 mg of nicotinic acid, 1.5 mg of thiamin hydrochloride, and 0.5 mg of biotin to 1 liter of water, and adjusting its pH to 7.2, then to which 30 g of calcium carbonate had been added) contained in a 2 1 buffle-attached Erlenmeyer flask and cultured therein at 30° C. for 12 to 16 hours. A total amount of the seed culturing medium was inoculated into 1,400 ml of a main culture medium (medium prepared by adding 60 g of glucose, 20 g of corn steep liquor, 25 g of ammonium chloride, 2.5 g of potassium dihydrogenphosphate, 0.75 g of magnesium sulfate heptahydrate, 50 mg of iron sulfate heptahydrate, 13 mg of manganese sulfate pentahydrate, 50 mg of calcium chloride, 6.3 mg of copper sulfate pentahydrate, 1.3 mg of zinc sulfate heptahydrate, 5 mg of nickel chloride hexahydrate, 1.3 mg of cobalt chloride hexahydrate, 1.3 mg of ammonium molybdenate tetrahydrate, 14 mg of nicotinic acid, 23 mg of β-alanine, 7 mg of thiamin hydrochloride, and 0.42 mg of biotin to 1 liter of water) contained in a 5 1 jar fermenter and cultured therein at 32° C., 1 vvm and 800 rpm while controlling the pH to 7.0 with aqueous ammonia. When glucose in the medium had been consumed, a glucose feeding solution (medium prepared by adding 400 g glucose and 45 g of ammonium chloride to 1 liter of water) was continuously added. The addition of feeding solution was carried out at a controlled speed so as to maintain the dissolved oxygen concentration within a range of 0.5 to 3 rpm After culturing for 29 hours, the culture was terminated. The cells were separated from the culture medium by centrifugation and then L-lysine hydrochloride in the supernatant was quantified by high performance liquid chromatography (HPLC). The results are shown in Table 2 below.
TABLE 2
|
|
StrainL-Lysine hydrochloride yield (g/l)
|
|
ATCC 130320
HD-18
No. 5845
No. 58pyc51
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As is apparent from the results shown in Table 2, the lysine productivity was improved by introducing the mutation, Val59Ala, in the hom gene or the mutation, Pro458Ser, in the pyc gene. Accordingly, it was found that the mutations are both effective mutations relating to the production of lysine. Strain, AHP-3, in which the mutation, Val59Ala, in the hom gene and the mutation, Pro458Ser, in the pyc gene have been introduced into the wild type ATCC 13032 strain together with the mutation, Thr331Ile in the lysC gene has been deposited on Dec. 5, 2000, in National Institute of Bioscience and Human Technology, Agency of Industrial Science and Technology (Higashi 1-1-3, Tsukuba-shi, Ibaraki, Japan) as FERM BP-7382.
EXAMPLE 3
Reconstruction of Lysine-Producing Strain Based on Genome Information
The lysine-producing mutant B-6 strain (Appl. Microbiol. Biotechnol., 32: 269-273 (1989)), which has been constructed by multiple round random mutagenesis with NTG and screening from the wild type ATCC 13032 strain, produces-a remarkably large amount of lysine hydrochloride when cultured in a jar at 32° C. using glucose as a carbon source. However, since the fermentation period is long, the production rate is less than 2.1 g/l/h. Breeding to reconstitute only effective mutations relating to the production of lysine among the estimated at least 300 mutations introduced into the B-6 strain in the wild type ATCC 13032 strain was performed.
(1) Identification of Mutation Point and Effective Mutation by Comparing the Gene Nucleotide Sequence of the B-6 Strain with that of the ATCC 13032 Strain
As described above, the nucleotide sequences of genes derived from the B-6 strain were compared with the corresponding nucleotide sequences of the ATCC 13032 strain genome represented by SEQ ID NOS:1 to 3501 and analyzed to identify many mutation points accumulated in the chromosome of the B-6 strain. Among these, a mutation, Val591Ala, in hom, a mutation, Thr311Ile, in lysC, a mutation, Pro458Ser, in pyc and a mutation, Ala213Thr, in zwf were specified as effective mutations relating to the production of lysine. Breeding to reconstitute the 4 mutations in the wild type strain and for constructing of an industrially important lysine-producing strain was carried out according to the method shown below.
(2) Construction of Plasmid for Gene Replacement Having Mutated Gene
The plasmid for gene replacement, pChom59, having the mutated hom gene and the plasmid for gene replacement, pCpyc458, having the mutated pyc gene were prepared in the above Example 2(2). Plasmids for gene replacement having the mutated lysC and zwf were produced as described below.
The lysC and zwf having mutation points were amplified by PCR, and inserted into a plasmid for gene replacement, pCES30, according to the TA cloning method described in Example 2(2) (Bio Experiment Illustrated, Vol. 3).
Separately, chromosomal DNA was prepared from the lysine-producing B-6 strain according to the above method of Saito et al. Using the chromosomal DNA as a template, PCR was carried out with Pfu turbo DNA polymerase (manufactured by Stratagene). In the mutated lysC gene, the DNAs having the nucleotide sequences represented by SEQ ID NOS:7006 and 7007 were used as the primer set. In the mutated zwf gene, the DNAs having the nucleotide sequences represented by SEQ ID NOS:7008 and 7009 as the primer set. The resulting PCR product was subjected to agarose gel electrophoresis-, and extracted and purified using GENEGLEAN Kit (manufactured by BIO 101). Then, the PCR product was allowed to react in the presence of Taq DNA polymerase (manufactured by Roche Diagnostics) and dATP at 72° C. for 10 minutes so that a nucleotide, adenine (A), was added to the 3′-end.
The above pCES30 T vector fragment and the mutated lysC gene (1.5 kb) or mutated zwf gene (2.3 kb) to which the nucleotide A had been added of the PCR product were concentrated by extraction with phenol/chloroform and precipitation with ethanol, and then ligated using Ligation Kit ver. 2. The ligation products were introduced into the ATCC 13032 strain according to the electroporation method, and cultured on BYG agar medium containing 25 μg/ml kanamycin at 30° C. for 2 days to obtain kanamycin-resistant transformants. Each of the resulting transformants was cultured overnight in BYG liquid medium containing 25 μg/ml kanamycin, and a plasmid was extracted from the culturing solution medium according to the alkali SDS method. As a result of digestion analysis using restriction enzymes, it was confirmed that the plasmid had a structure in which the 1.5 kb or 2.3 kb DNA fragment had been inserted into pCES30. The plasmids thus constructed were named respectively pClysC311 and pCzwf213.
(3) Introduction of Mutation, Thr311Ile, in lysC into One Point Mutant HD-1
Since the one mutation point mutant HD-1 in which the mutation, Val59Ala, in hom was introduced into the wild type ATCC 13032 strain had been obtained in Example 2(2), the mutation, Thr311Ile, in lysC was introduced into the HD-1 strain using pClysC311 produced in the above (2) according to the gene replacement method described in Example 2(2). PCR was carried out using chromosomal DNA of the resulting strain and, as the primer set, DNAs having the nucleotide sequences represented by SEQ ID NOS:7006 and 7007 in the same manner as in Example 2(2). As a result of the fact that the nucleotide sequence of the PCR product was determined in the usual manner, it was confirmed that the strain which was named AHD-2 was a two point mutant having the mutated lysC gene in addition to the mutated hom gene.
(4) Introduction of Mutation, Pro458Ser, in pyc into Two Point Mutant AHD-2
The mutation, Pro458Ser, in pyc was introduced into the AHD-2 strain using the pCpyc458 produced in Example 2(2) by the gene replacement method described in Example 2 (2). PCR was carried out using chromosomal DNA of the resulting strain and, as the primer set, DNAs having the nucleotide sequences represented by SEQ ID NOS:7004 and 7005 in the same manner as in Example 2(2). As a result of the fact that the nucleotide sequence of the PCR product was determined in the usual manner, it was confirmed that the strain which was named AHD-3 was a three point mutant having the mutated pyc gene in addition to the mutated hom gene and lysC gene.
(5) Introduction of Mutation, Ala213Thr, in zwf into Three Point Mutant AHP-3
The mutation, Ala213Thr, in zwf was introduced into the AHP-3 strain using the pCzwf458 produced in the above (2) by the gene replacement method described in Example 2(2). PCR was carried out using chromosomal DNA of the resulting strain and, as the primer set, DNAs having the nucleotide sequences represented by SEQ ID NOS:7008 and 7009 in the same manner as in Example 2(2). As a result of the fact that the nucleotide sequence of the PCR product was determined in the usual manner, it was confirmed that the strain which was named APZ-4 was a four point mutant having the mutated zwf gene in addition to the mutated hom gene, lysC gene and pyc gene.
(6) Lysine Production Test on HD-1, AHD-2, AHP-3 and APZ-4 Strains
The HD-1, AHD-2, AHP-3 and APZ-4 strains obtained above were subjected to a culture test in a 5 1 jar fermenter in accordance with the method of Example 2(3).
Table 3 shows the results.
TABLE 3
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L-Lysine hydrochlorideProductivity
Strain(g/l)(g/l/h)
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HD-180.3
AHD-2732.5
AHP-3802.8
APZ-4863.0
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Since the lysine-producing mutant B-6 strain which has been bred based on the random mutation and selection shows a productivity of less than 2.1 g/l/h, the APZ-4 strain showing a high productivity of 3.0 g/l/h is useful in industry.
(7) Lysine Fermentation by APZ-4 Strain at High Temperature
The APZ-4 strain, which had been reconstructed by introducing 4 effective mutations into the wild type strain, was subjected to the culturing test in a 5 l jar fermenter in the same manner as in Example 2(3), except that the culturing temperature was changed to 40° C.
The results are shown in Table 4.
TABLE 4
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TemperatureL-Lysine hydrochlorideProductivity
(° C.)(g/l)(g/l/h)
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32863.0
40953.3
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As is apparent from the results shown in Table 4, the lysine hydrochloride titer and productivity in culturing at a high temperature of 40° C. comparable to those at 32° C. were obtained. In the mutated and bred lysine-producing B-6 strain constructed by repeating random mutation and selection, the growth and the lysine productivity are lowered at temperatures exceeding 34° C. so that lysine fermentation cannot be carried out, whereas lysine fermentation can be carried out using the APZ-4 strain at a high temperature of 40° C. so that the load of cooling is greatly reduced and it is industrially useful. The lysine fermentation at high temperatures can be achieved by reflecting the high temperature adaptability inherently possessed by the wild type strain on the APZ-4 strain.
As demonstrated in the reconstruction of the lysine-producing strain, the present invention provides a novel breeding method effective for eliminating the problems in the conventional mutants and acquiring industrially advantageous strains. This methodology which reconstitutes the production strain by reconstituting the effective mutation is an approach which is efficiently carried out using the nucleotide sequence information of the genome disclosed in the present invention, and its effectiveness was found for the first time in the present invention.
EXAMPLE 4
Production of DNA Microarray and use thereof
A DNA microarray was produced based on the nucleotide sequence information of the ORF deduced from the full nucleotide sequences of Corynebacterium glutamicum ATCC 13032 using software, and genes of which expression is fluctuated depending on the carbon source during culturing were searched.
(1) Production of DNA Microarray
Chromosomal DNA was prepared from Corynebacterium glutamicum ATCC 13032 by the method of Saito et al. (Biochem. Biophys. Acta, 72: 619 (1963)). Based on 24 genes having the nucleotide sequences represented by SEQ ID NOS:207, 3433, 281, 3435, 3439, 765, 3445, 1226, 1229, 3448, 3451, 3453, 3455, 1743, 3470, 2132, 3476, 3477, 3485, 3488, 3489, 3494, 3496, and 3497 from the ORFs shown in Table 1 deduced from the full genome nucleotide sequence of Corynebacterium glutamicum ATCC 13032 using software and the nucleotide sequence of rabbit globin gene (GenBank Accession No. V00882) used as an internal standard, oligo DNA primers for PCR amplification represented by SEQ ID NOS:7010 to 7059 targeting the nucleotide sequences of the genes were synthesized in a usual manner.
As the oligo DNA primers used for the PCR,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7010 and 7011 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:207,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7012 and 7013 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3433,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7014 and 7015 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:281,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7016 and 7017 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3435,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7018 and 7019 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3439,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7020 and 7021 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:765,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7022 and 7023 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3445,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7024 and 7025 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:1226,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7026 and 7027 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:1229,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7028 and 7029 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3448,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7030 and 7031 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3451,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7032 and 7033 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3453,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7034 and 7035 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3455,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7036 and 7037 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:1743,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7038 and 7039 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3470,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7040 and 7041 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:2132,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7042 and 7043 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3476,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7044 and 7045 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3477,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7046 and 7047 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3485,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7048 and 7049 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3488,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7050 and 7051 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3489,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7052 and 7053 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3494,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7054 and 7055 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3496,
DNAs having the nucleotide sequence represented by SEQ ID NOS:7056 and 7057 were used for the amplification of the DNA having the nucleotide sequence represented by SEQ ID NO:3497, and
DNAs having the nucleotide sequence represented by SEQ ID NOS:7058 and 7059 were used for the amplification of the DNA having the nucleotide sequence of the rabbit globin gene,
as the respective primer set.
The PCR was carried for 30 cycles with each cycle consisting of 15 seconds at 95° C. and 3 minutes at 68° C. using a thermal cycler (GeneAmp PCR system 9600, manufactured by Perkin Elmer), TaKaRa EX-Taq (manufactured by Takara Shuzo), 100 ng of the chromosomal DNA and the buffer attached to the TaKaRa Ex-Taq reagent. In the case of the rabbit globin gene, a single-stranded cDNA which had been synthesized from rabbit globin mRNA (manufactured by Life Technologies) according to the manufacture's instructions using a reverse transcriptase RAV-2 (manufactured by Takara Shuzo). The PCR product of each gene thus amplified was subjected to agarose gel electrophoresis and extracted and purified using QIAquick Gel Extraction Kit (manufactured by QIAGEN). The purified PCR product was concentrated by precipitating it with ethanol and adjusted to a concentration of 200 ng/μl. Each PCR product was spotted on a slide glass plate (manufactured by Matsunami Glass) having MAS coating in 2 runs using GTMASS SYSTEM (manufactured by Nippon Laser & Electronics Lab.) according to the manufacture's instructions.
(2) Synthesis of Fluorescence Labeled cDNA
The ATCC 13032 strain was spread on BY agar medium (medium prepared by adding 20 g of peptone (manufactured by Kyokuto Pharmaceutical), 5 g of yeast extract (manufactured by Difco), and 16 g of Bactoagar (manufactured by Difco) to in 1 liter of water and adjusting its pH to 7.2) and cultured at 30° C. for 2 days. Then, the cultured strain was further inoculated into 5 ml of BY liquid medium and cultured at 30° C. overnight. Then, the cultured strain was further inoculated into 30 ml of a minimum medium (medium prepared by adding 5 g of ammonium sulfate, 5 g of urea, 0.5 g of monopotassium dihydrogenphosphate, 0.5 g of dipotassium monohydrogenphosphate, 20.9 g of morpholinopropanesulfonic acid, 0.25 g of magnesium sulfate heptahydrate, 10 mg of calcium chloride dihydrate, 10 mg of manganese sulfate monohydrate, 10 mg of ferrous sulfate heptahydrate, 1 mg of zinc sulfate heptahydrate, 0.2 mg copper sulfate, and 0.2 mg biotin to 1 liter of water, and adjusting its pH to 6.5) containing 110 mmol/l glucose or 200 mmol/l ammonium acetate, and cultured in an Erlenmyer flask at 30° to give 1.0 of absorbance at 660 nm. After the cells were prepared by centrifuging at 4° C. and 5,000 rpm for 10 minutes, total RNA was prepared from the resulting cells according to the method of Bormann et al. (Molecular Microbiology, 6: 317-326 (1992)). To avoid contamination with DNA, the RNA was treated with DnaseI (manufactured by Takara Shuzo) at 37° C. for 30 minutes and then further purified using Qiagen RNeasy MiniKit (manufactured by QIAGEN) according to the manufacture's instructions. To 30 μg of the resulting total RNA, 0.6 μl of rabbit globin mRNA (50 ng/μl, manufactured by Life Technologies) and 1 μl of a random 6 mer primer (500 ng/μl, manufactured by Takara Shuzo) were added for denaturing at 65° C. for 10 minutes, followed by quenching on ice. To the resulting solution, 6 Al of a buffer attached to SuperScript II (manufactured by Lifetechnologies), 3 μl of 0.1 mol/l DTT, 1.5 μl of dNTPs (25 mmol/l dATP, 25 mmol/l dCTP, 25 mmol/l dGTP, 10 mmol/l dTTP), 1.5 μl of Cy5-dUTP or Cy3-dUTP (manufactured by NEN) and 2 μl of SuperScript II were added, and allowed to stand at 25° C. for 10 minutes and then at 42° C. for 110 minutes. The RNA extracted from the cells using glucose as the carbon source and the RNA extracted from the cells using ammonium acetate were labeled with Cy5-dUTP and Cy3-dUTP, respectively. After the fluorescence labeling reaction, the RNA was digested by adding 1.5 μl of 1 mol/l sodium hydroxide-20 mmol/l EDTA solution and 3.0 μl of 10% SDS solution, and allowed to stand at 65° C. for 10 minutes. The two cDNA solutions after the labeling were mixed and purified using Qiagen PCR purification Kit (manufactured by QIAGEN) according to the manufacture's instructions to give a volume of 10 μl.
(3) Hybridization
UltraHyb (110 μl) (manufactured by Ambion) and the fluorescence-labeled cDNA solution (10 μl) were mixed and subjected to hybridization and the subsequent washing of slide glass using GeneTAC Hybridization Station (manufactured by Genomic Solutions) according to the manufacture's instructions. The hybridization was carried out at 50° C., and the washing was carried out at 25° C.
(4) Fluorescence Analysis
The fluorescence amount of each DNA array having the fluorescent cDNA hybridized therewith was measured using ScanArray 4000 (manufactured by GSI Lumonics).
Table 5 shows the Cy3 and Cy5 signal intensities of the genes having been corrected on the basis of the data of the rabbit globin used as the internal standard and the Cy3/Cy5 ratios.
TABLE 5
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SEQ ID NOCy3 intensityCy5 intensityCy3/Cy5
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207524832401.62
3433223926940.83
281237025950.91
3435256625151.02
3439559769440.81
765613449431.24
3455116912840.91
1226130114930.87
1229116811311.03
3448118715940.74
3451284538590.74
3453349817052.05
3455149111441.30
1743197218411.07
3470475237641.26
2132117310851.08
3476184714201.30
3477128411641.10
3485453980140.57
348834289139824.52
348943645149729.16
3494319925031.28
3496342823641.45
3497384833581.15
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The ORF function data estimated by using software were searched for SEQ ID NOS:3488 and 3489 showing remarkably strong Cy3 signals. As a result, it was found that SEQ ID NOS:3488 and 3489 are a maleate synthase gene and an isocitrate lyase gene, respectively. It is known that these genes are transcriptionally induced by acetic acid in Corynebacterium glutamicum (Archives of Microbiology, 168: 262-269 (1997)).
As described above, a gene of which expression is fluctuates could be discovered by synthesizing appropriate oligo DNA primers based on the ORF nucleotide sequence information deduced from the full genomic nucleotide sequence information of Corynebacterium glutamicum ATCC 13032 using software, amplifying the nucleotide sequences of the gene using the genome DNA of Corynebacterium glutamicum as a template in the PCR reaction, and thus producing and using a DNA microarray.
This Example shows that the expression amount can be analyzed using a DNA microarray in the 24 genes. On the other hand, the present DNA microarray techniques make it possible to prepare DNA microarrays having thereon several thousand gene probes at once. Accordingly, it is also possible to prepare DNA microarrays having thereon all of the ORF gene probes deduced from the full genomic nucleotide sequence of Corynebacterium glutamicum ATCC 13032 determined by the present invention, and analyze the expression profile at the total gene level of Corynebacterium glutamicum using these arrays.
EXAMPLE 5
Homology Search Using Corynebacterium glutamicum Genome Sequence
(1) Search of Adenosine Deaminase
The amino acid sequence (ADD_ECOLI) of Escherichia coli adenosine deaminase was obtained from Swiss-prot Database as the amino acid sequence of the protein of which function had been confirmed as adenosine deaminase (EC3.5.4.4). By using the full length of this amino acid sequence as a query, a homology search was carried out on a nucleotide sequence database of the genome sequence of Corynebacterium glutamicum or a database of the amino acids in the ORF region deduced from the genome sequence using FASTA program (Proc. Natl. Acad. Sci. ISA, 85: 2444-2448 (1988)). A case where E-value was le−10 or less was judged as being significantly homologous. As a result, no sequence significantly homologous with the Escherichia coli adenosine deaminase was found in the nucleotide sequence database of the genome sequence of Corynebacterium glutamicum or the database of the amino acid sequences in the ORF region deduced from the genome sequence. Based on these results, it is assumed that Corynebacterium glutamicum contains no ORF having adenosine deaminase activity and thus has no activity of converting adenosine into inosine.
(2) Search of Glycine Cleavage Enzyme
The sequences (GCSP_ECOLI, GCST_ECOLI and GCSH_ECOTI) of glycine decarboxylase, aminomethyl transferase and an aminomethyl group carrier each of which is a component of Escherichia coli glycine cleavage enzyme as the amino acid sequence of the protein, of which function had been confirmed as glycine cleavage enzyme (EC2.1.2.10), were obtained from Swiss-prot Database.
By using these full-length amino acid sequences as a query, a homology search was carried out on a nucleotide sequence database of the genome sequence of Corynebacterium glutamicum or a database of the ORF amino acid sequences deduced from the genome sequence using FASTA program. A case where E-value was le−10 or less was judged as being significantly homologous. As a result, no sequence significantly homologous with the glycine decarboxylase, the aminomethyl transferase or the aminomethyl group carrier each of which is a component of Escherichia coli glycine cleavage enzyme, was found in the nucleotide sequence database of the genome sequence of Corynebacterium glutamicum or the database of the ORF amino acid sequences estimated from the genome sequence. Based on these results, it is assumed that Corynebacterium glutamicum contains no ORF having the activity of glycine decarboxylase, aminomethyl transferase or the aminomethyl group carrier and thus has no activity of the glycine cleavage enzyme.
(3) Search of IMP Dehydrogenase
The amino acid sequence (IMDH ECOLI) of Escherichia coli IMP dehydrogenase as the amino acid sequence of the protein, of which function had been confirmed as IMP dehydrogenase (EC1.1.1.205), was obtained from Swiss-prot Database. By using the full length of this amino acid sequence as a query, a homology search was carried out on a nucleotide sequence database of the genome sequence of Corynebacterium glutamicum or a database of the ORF amino acid sequences predicted from the genome sequence using FASTA program. A case where E-value was le−10 or less was judged as being significantly homologous. As a result, the amino acid sequences encoded by two ORFs, namely, an ORF positioned in the region of the nucleotide sequence No. 615336 to 616853 (or ORF having the nucleotide sequence represented by SEQ ID NO:672) and another ORF positioned in the region of the nucleotide sequence No. 616973 to 618094 (or ORF having the nucleotide sequence represented by SEQ ID NO:674) were significantly homologous with the ORFs of Escherichia coli IMP dehydrogenase. By using the above-described predicted amino acid sequence as a query in order to examine the similarity of the amino acid sequences encoded by the ORFs with IMP dehydrogenases of other organisms in greater detail, a search was carried out on GenBank (http://www.ncbi.nlm.nih.gov/) nr-aa database (amino acid sequence database constructed on the basis of GenBankCDS translation products, PDB database, Swiss-Prot database, PIR database, PRF database by eliminating duplicated registrations) using BLAST program. As a result, both of the two amino acid sequences showed significant homologies with IMP dehdyrogenases of other organisms and clearly higher homologies with IMP dehdyrogenases than with amino acid sequences of other proteins, and thus, it was assumed that the two ORFs would function as IMP dehydrogenase. Based on these results, it was therefore assumed that Corynebacterium glutamicum has two ORFs having the IMP dehydrogenase activity.
EXAMPLE 6
Proteome Analysis of Proteins Derived from Corynebacterium glutamicum
(1) Preparations of Proteins Derived from Corynebacterium glutamicum ATCC 13032, FERM BP-7134 and FERM BP-158
Culturing tests of Corynebacterium glutamicum ATCC 13032 (wild type strain), Corynebacterium glutamicum FERM BP-7134 (lysine-producing strain) and Corynebacterium glutamicum (FERM BP-158, lysine-highly producing strain) were carried out in a 5 1 jar fermenter according to the method in Example 2(3). The results are shown in Table 6.
TABLE 6
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StrainL-Lysine yield (g/l)
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ATCC 130320
FERM BP-713445
FERM BP-15860
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After culturing, cells of each strain were recovered by centrifugation. These cells were washed with Tris-HCl buffer (10 mmol/l Tris-HCl, pH 6.5, 1.6 mg/ml protease inhibitor (COMPLETE; manufactured by Boehringer Mannheim)) three times to give washed cells which could be stored under freezing at −80° C. The freeze-stored cells were thawed before use, and used as washed cells.
The washed cells described above were suspended in a disruption buffer (10 mmol/l Tris-HCl, pH 7.4, 5 mmol/l magnesium chloride, 50 mg/l RNase, 1.6 mg/ml protease inhibitor (COMPLETE: manufactured by Boehringer Mannheim)), and disrupted with a disrupter (manufactured by Brown) under cooling. To the resulting disruption solution, DNase was added to give a concentration of 50 mg/l, and allowed to stand on ice for 10 minutes. The solution was centrifuged (5,000×g, 15 minutes, 4° C.) to remove the undisrupted cells as the precipitate, and the supernatant was recovered.
To the supernatant, urea was added to give a concentration of 9 mol/l, and an equivalent amount of a lysis buffer (9.5 mol/l urea, 2% NP-40, 2% Ampholine, 5% mercaptoethanol, 1.6 mg/ml protease inhibitor (COMPLETE; manufactured by Boehringer Mannheim) was added thereto, followed by thoroughly stirring at room temperature for dissolving.
After being dissolved, the solution was centrifuged at 12,000×g for 15 minutes, and the supernatant was recovered.
To the supernatant, ammonium sulfate was added to the extent of 80% saturation, followed by thoroughly stirring for dissolving.
After being dissolved, the solution was centrifuged (16,000×g, 20 minutes, 4° C.), and the precipitate was recovered. This precipitate was dissolved in the lysis buffer again and used in the subsequent procedures as a protein sample. The protein concentration of this sample was determined by the method for quantifying protein of Bradford.
(2) Separation of Protein by Two Dimensional Electrophoresis
The first dimensional electrophoresis was carried out as described below by the isoelectric electrophoresis method.
A molded dry IPG strip gel (pH 4-7, 13 cm, Immobiline DryStrips; manufactured by Amersham Pharmacia Biotech) was set in an electrophoretic apparatus (Multiphor II or IPGphor; manufactured by Amersham Pharmacia Biotech) and a swelling solution (8 mol/l urea, 0.5% Triton X-100, 0.69 dithiothreitol, 0:5% Ampholine, pH 3-10) was packed therein, and the gel was allowed to stand for swelling 12 to 16 hours.
The protein sample prepared above was dissolved in a sample solution (9 mol/l urea, 2% CRAPS, 1% dithiothreitol, 2t Ampholine, pH 3-10), and then about 100 to 500 )g (in terms of protein) portions thereof were taken and added to the swollen IPG strip gel.
The electrophoresis was carried out in the 4 steps as defined below under controlling the temperature to 20° C.:
- step 1: 1 hour under a gradient mode of 0 to 500V;
- step 2: 1 hour under a gradient mode of 500 to 1,000 V;
- step 3: 4 hours under a gradient mode of 1,000 to 8,000 V; and
- step 4: 1 hour at a constant voltage of 8,000 V.
After the isoelectric electrophoresis, the IPG strip gel was put off from the holder and soaked in an equilibration buffer A (50 mmol/l Tris-HCl, pH 6.8, 30% glycerol, 1% SDS, 0.254 dithiothreitol) for 15 minutes and another equilibration buffer B (50 mmol/l Tris-HCl, pH 6.8, 6 mol/l urea, 30% glycerol, 1% SOS, 0.45% iodo acetami de) for 15 minutes to sufficiently equilibrate the gel.
After the equilibrium, the IPG strip gel was lightly rinsed in an SDS electrophoresis buffer (1.4% glycine, 0.1% SDS, 0.3% Tris-HCl, pH 8.5), and the second dimensional elect-ophoresis depending on molecular weight was carried out as described below to separate the proteins.
Specifically, the above IPG strip gel was closely placed on 14% polyacrylamide slub gel (14% polyacrylamide, 0.37% bisacrylamide, 37.5 mmol/l Tris-HCl, pH 8.8, 0.1% SDS, 0.1% TEMED, 0.1% ammonium persulfate) and subjected to electrophoresis under a constant voltage of 30 mA at 20° C. for 3 hours to separate the proteins.
(3) Detection of Protein Spot
Coomassie staining was performed by the method of Gorg et al. (Electrophoresis, 9: 531-546 (1988)) for the slub gel after the second dimensional electrophoresis. Specifically, the slub gel was stained under shaking at 25° C. for about 3 hours, the excessive coloration was removed with a decoloring solution, and the gel was thoroughly washed with distilled water.
The results are shown in FIG. 2. The proteins derived from the ATCC 13032 strain (FIG. 2A), FERM BP-7134 strain (FIG. 2B) and FERM BP-158 strain (FIG. 2C) could be separated and detected as spots.
(4) In-gel Digestion of Detected Protein Spot
The detected spots were each cut out from the gel and transferred into siliconized tube, and 400 μl of 100 mmol/l ammonium bicarbonate : acetonitrile solution (1:1, v/v) was added thereto, followed by shaking overnight and freeze-dried as such. To the dried gel, 10 μl of a lysylendopeptidase (LysC) solution (manufactured by WAKO, prepared with 0.1% SDS-containing 50 mmol/l ammonium bicarbonate to give a concentration of 100 ng/μl) was added and the gel was allowed to stand for swelling at 0° C. for 45 minutes, and then allowed to stand at 37° C. for 16 hours. After removing the LysC solution, 20 μl of an extracting solution (a mixture of 60% acetonitrile and 5% formic acid) was added, followed by ultrasonication at room temperature for 5 minutes to disrupt the gel. After the disruption, the extract was recovered by centrifugation (12,000 rpm, 5 minutes, room temperature). This operation was repeated twice to recover the whole extract. The recovered extract was concentrated by centrifugation in vacuo to halve the liquid volume. To the concentrate, 20 μl of 0.1% trifluoroacetic acid was added, followed by thoroughly stirring, and the mixture was subjected to desalting using ZipTip (manufactured by Millipore). The protein absorbed on the carriers of ZipTip was eluted with 5 μl of α-cyano-4-hydroxycinnamic acid for use as a sample solution for analysis.
(5) Mass Spectrometry and Amino Acid Sequence Analysis of Protein Spot with Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometer (MALDI-TOFMS)
The sample solution for analysis was mixed in the equivalent amount with a solution of a peptide mixture for mass calibration (300 nmol/l Angiotensin II, 300 nmol/l Neurotensin, 150 nmol/l ACTHclip 18-39, 2.3 μmol/l bovine insulin B chain), and 1 μl of the obtained solution was spotted on a stainless probe and crystallized by spontaneously drying.
As measurement instruments, REFLEX MALDI-TOF mass spectrometer (manufactured by Bruker) and an N2 laser (337 nm) were used in combination.
The analysis by PMF (peptide-mass finger printing) was carried out using integration spectra data obtained by measuring 30 times at an accelerated voltage of 19.0 kV and a detector voltage of 1.50 kV under reflector mode conditions. Mass calibration was carried out by the internal standard method.
The PSD (post-source decay) analysis was carried out using integration spectra obtained by successively altering the reflection voltage and the detector voltage at an accelerated voltage of 27.5 kV.
The masses and amino acid sequences of the peptide fragments derived from the protein spot after digestion were thus determined.
(6) Identification of Protein Spot
From the amino acid sequence information of the digested peptide fragments derived from the protein spot obtained in the above (5), ORFs corresponding to the protein were searched on the genome sequence database of Corynebacterium glutamicum ATCC 13032 as constructed in Example 1 to identify the protein.
The identification of the protein was carried out using MS-Fit program and MS-Tag program of intranet protein prospector.
(a) Search and Identification of Gene Encoding High-Expression Protein
In the proteins derived from Corynebacterium glutamic ATCC 13032 showing high expression amounts in CBB-staining shown in FIG. 2A, the proteins corresponding to Spots-1, 2, 3, 4 and 5 were identified by the above method.
As a result, it was found that Spot-i corresponded to enolase which was a protein having the amino acid sequence of SEQ ID NO:4585; Spot-2 corresponded to phosphoglycelate kinase which was a protein having the amino acid sequence of SEQ ID NO:5254; Spot-3 corresponded to glyceraldehyde-3-phosphate dehydrogenase which was a protein having the amino acid sequence represented by SEQ ID NO:5255; Spot-4 corresponded to fructose bis-phosphate aldolase which was a protein having the amino acid sequence represented by SEQ ID NO:6543; and Spot-5 corresponded to triose phosphate isomerase which was a protein having the amino acid sequence represented by SEQ ID NO:5252.
These genes, represented by SEQ ID NOS:1085, 1754, 1775, 3043 and 1752 encoding the proteins corresponding to Spots-1, 2, 3, 4 and 5, respectively, encoding the known proteins are important in the central metabolic pathway for maintaining the life of the microorganism. Particularly, it is suggested that the genes of Spots-2, 3 and 5 form an operon and a high-expression promoter is encoded in the upstream thereof (J. of Bacteriol., 174: 6067-6086 (1992)).
Also, the protein corresponding to Spot-9 in FIG. 2 was identified in the same manner as described above, and it was found that Spot-9 was an elongation factor Tu which was a protein having the amino acid sequence represented by SEQ ID NO:6937, and that, the protein was encoded by DNA having the nucleotide sequence represented by SEQ ID NO:3437.
Based on these results, the proteins having high expression level were identified by proteome analysis using the genome sequence database of Corynebacterium glutamicum constructed in Example 1. Thus, the nucleotide sequences of the genes encoding the proteins and the nucleotide sequences upstream thereof could be searched simultaneously Accordingly, it is shown that nucleotide sequences having a function as a high-expression promoter can be efficiently selected.
(b) Search and Identification of Modified Protein
Among the proteins derived from Corynebacterium glutamicum FERM BP-7134 shown in FIG. 2E, Spots-6, 7 and 8 were identified by the above method. As a result, these three spots all corresponded to catalase which was a protein having the amino acid sequence represented by SEQ ID NO:3785.
Accordingly, all of Spots-6, 7 and 8 detected as spots differing in isoelectric mobility were all products derived from a catalase gene having the nucleotide sequence represented by SEQ ID NO:285. Accordingly, it is shown that the catalase derived from Corynebacterium glutamicum FERM BP-7134 was modified after the translation.
Based on these results, it is confirmed that various modified proteins can be efficiently searched by proteome analysis using the genome sequence database of Corynebacterium glutamicum constructed in Example 1.
(c) Search and Identification of Expressed Protein Effective in Lysine Production
It was found out that in FIG. 2A (ATCC 13032: wild type strain), FIG. 2B (FERM BP-7134: lysine-producing strain) and FIG. 2C (FERM BP-158: lysine-highly producing strain), the catalase corresponding to Spot-8 and the elongation factor Tu corresponding to Spot-9 as identified above showed the higher expression level with an increase in the lysine productivity.
Based on these results, it was found that hopeful mutated proteins can be efficiently searched and identified in breeding aiming at strengthening the productivity of a target product by the proteome analysis using the genome sequence database of Corynebacterium glutamicum constructed in Example 1.
Moreover, useful mutation points of useful mutants can be easily specified by searching the nucleotide sequences (nucleotide sequences of promoter, ORF, or the like) relating to the identified proteins, using the above database and using primers designed on the basis of the sequences. As a result of the fact that the mutation points are specified, industrially useful mutants which have the useful mutations or other useful mutations derived therefrom can be easily bred.
While the invention has been described in detail and with reference to specific embodiments thereof, it will be apparent to one of skill in the art that various changes and modifications can be made therein without departing from the spirit and scope thereof. All references cited herein are incorporated in their entirety.
Patent Application
20060228712
