Compositions and methods for fumonisin detoxification

Abstract

Compositions and methods for the complete detoxification of fumonisin and fumonisin degradation products are provided. Particularly, nucleotide sequences corresponding to the detoxification enzymes are provided. The sequences find use in preparing expression cassettes for the transformation of a broad variety of host cells and organisms.

Description

FIELD OF THE INVENTION

The invention relates to compositions and methods for detoxification or degradation of fumonisins and related toxins. The method has broad application in agricultural biotechnology and crop agriculture and in the improvement of food grain quality.

BACKGROUND OF THE INVENTION

Fungal diseases are common problems in crop agriculture. Many strides have been made against plant diseases as exemplified by the use of hybrid plants, pesticides, and improved agricultural practices. However, as any grower or home gardener can attest, the problems of fungal plant disease continue to cause difficulties in plant cultivation. Thus, there is a continuing need for new methods and materials for solving the problems caused by fungal diseases of plants.

These problems can be met through a variety of approaches. For example, the infectious organisms can be controlled through the use of agents that are selectively biocidal for the pathogens. Another method is interference with the mechanism by which the pathogen invades the host crop plant. Yet another method, in the case of pathogens that cause crop losses, is interference with the mechanism by which the pathogen causes injury to the host crop plant. In the case of pathogens that produce toxins that are undesirable to mammals or other animals that feed on the crop plants, interference with toxin production, storage, or activity can be beneficial.

Since their discovery and structural elucidation in 1988 (Bezuidenhout et al. (1988) Journal Chem. Soc., Chem. Commun. 1988:743-745), fumonisins have been recognized as a potentially serious problem in maize-fed livestock. They are linked to several animal toxicoses including leukoencephalomalacia (Marasas et al. (1988) Onderstepoort J. Vet. Res. 55:197-204; Wilson et al. (1990) American Association of Veterinary Laboratory Diagnosticians: Abstracts 33 rd Annual Meeting, Denver, Colo., Madison, Wis., USA) and porcine pulmonary edema (Colvin et al. (1992) Mycopathologia 117:79-82). Fumonisins are also suspected carcinogens (Geary et al. (1971) Coord. Chem. Rev. 7:81; Gelderblom et al. (1991) Carcinogenesis 12:1247-1251; Gelderblom et al. (1992) Carcinogenesis 13:433-437). Fusarium isolates in section Liseola produce fumonisins in culture at levels from 2 to >4000 ppm (Leslie et al. (1992) Phytopathology 82:341-345). Isolates from maize (predominantly mating population A) are among the highest producers of fumonisin (Leslie et al., supra). Fumonisin levels detected in field-grown maize have fluctuated widely depending on location and growing season, but both preharvest and post-harvest surveys of field maize have indicated that the potential for high levels of fumonisins exists (Murphy et al. (1993) J. Agr. Food Chem. 41:263-266). Surveys of food and feed products have also detected fumonisin (Holcomb et al. (1993) J. Agr. Food Chem. 41:764-767; Hopmans et al. (1993) J. Agr. Food Chem. 41:1655-1658); Sydenham et al. (1991) J. Agr. Food Chem. 39:2014-2018). The etiology of Fusarium ear mold is poorly understood, although physical damage to the ear and certain environmental conditions can contribute to its occurrence (Nelson et al. (1992) Mycopathologia 117:29-36). Fusarium can be isolated from most field grown maize, even when no visible mold is present. The relationship between seedling infection and stalk and ear diseases caused by Fusarium is not clear. Genetic resistance to visible kernel mold has been identified (Gendloff et al. (1986) Phytopathology 76:684-688; Holley et al. (1989) Plant Dis. 73:578-580), but the relationship between visible mold and fumonisin production has yet to be elucidated.

Fumonisins have been shown in in vitro mammalian cell studies to inhibit sphingolipid biosynthesis through inhibition of the enzyme sphingosine N-acetyl transferase, resulting in the accumulation of the precursor sphinganine (Norred et al. (1992) Mycopathologia 117:73-78; Wang et al. (1991) Biol. Chem. 266:14486; Yoo et al. (1992) Toxicol. Appl. Pharmacol. 114:9-15; Nelson et al. (1993) Annu. Rev. Phytpathol. 31:233-252). It is likely that inhibition of this pathway accounts for at least some of fumonisin's toxicity, and support for this comes from measures of sphinganine:sphingosine ratios in animals fed purified fumonisin (Wang et al. (1992) J. Nutr. 122:1706-1716). Fumonisins also affect plant cell growth (Abbas et al. (1992) Weed Technol. 6:548-552; Van Asch et al. (1992) Phytopathology 82:1330-1332; Vesonder et al. (1992) Arch. Environ. Contam. Toxicol. 23:464-467). Kuti et al. (1993) (Abstract, Annual Meeting American Phytopathological Society, Memphis, Tenn.: APS Press) reported on the ability of exogenously added fumonisins to accelerate disease development and increase sporulation of Fusarium moniliform and F. oxysporum on tomato.

Enzymes that degrade the fungal toxin fumonisin to the compound AP1 have been identified in U.S. Pat. No. 5,716,820 and pending U.S. patent application Ser. Nos. 08/888,949 and 08/888,950, both filed Jul. 7, 1997, and hereby incorporated by reference. Plants expressing a fumonisin esterase enzyme, infected by fumonisin producing fungus, and tested for fumonisin and AP1 were found to have low levels of fumonisin but high levels of AP1. AP1 is less toxic than fumonisin to plants and probably also animals, but contamination with AP1 is still a concern. The best result would be complete detoxification of fumonisin to a non-toxic form. Therefore additional enzymes capable of degrading fumonisin and fumonisin catabolic products are necessary for the complete detoxification of fumonisin.

SUMMARY OF THE INVENTION

A fumonisin detoxification gene cluster is provided. Particularly, the nucleotide sequence for the cluster from Bacterium 2412.1 (American Type Culture Collection Deposit Number 55552) is provided. At least twelve structural genes and three regulatory gene are provided. The sequences represent a catabolic pathway for fumonisins and fumonisin-like compounds.

Compositions and methods for catabolism and detoxification of fumonisin, fumonisin-related toxins, and fumonisin-degradation products are provided. In particular, proteins involved in catabolism and transmembrane transport of fumonisin and fumonisin catabolic products are provided. Nucleotide sequences corresponding to the proteins are also included. The compositions are useful in the complete detoxification and degradation of fumonisin. The nucleotide sequences can be used in expression cassettes for transformation of host cells of interest. The compositions and methods of the invention provide a catabolic pathway for fumonisin. Thus, organisms can be genetically modified to provide for the complete catabolism and detoxification of fumonisin and fumonisin-related toxins.

In particular, expression cassettes for expression of the sequences in plants and other organisms are provided as well as transformed plants and other host cells.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 sets forth the proposed pathway for fumonisin degradation by Bacterium 2412.1.

FIG. 2 illustrates the overlap of the individual lambda clones and cosmids which were sequenced to reconstruct the fumonisin catabolic gene cluster.

FIG. 3 provides a map of the open reading frames in the fumonisin catabolic gene cluster of bacterium 2412.1

FIG. 4 schematically illustrates a plasmid vector comprising the gene for one of the fumonisin degradative enzymes of the invention operably linked to the ubiquitin promoter.

DETAILED DESCRIPTION OF THE INVENTION

A catabolic pathway for detoxification and degradation of fumonisin is provided. Particularly, enzymes involved in the degradation of fumonisin from Bacterium 2412.1 and nucleotide sequences encoding such enzymes are disclosed. Such enzymes and nucleotide sequences find use in the breakdown of fumonisin as well as fumonisin-degradation products. In this regard, enzymes can be synthesized and utilized or, alternatively, organisms can be transformed with the DNA sequences of the invention and used to detoxify fumonisin.

A proposed pathway for the degradation of fumonisin by Bacterium 2412.1 is provided in FIG. 1 . The present invention provides enzymes and nucleotide sequences encoding the enzymes involved in this degradation pathway for fumonisin. In particular, the present invention provides for isolated nucleic acid molecules comprising the fumonisin catabolic gene cluster for Bacterium 2412.1 set forth in SEQ ID NO: 1, or the DNA sequences obtained from the overlapping clones deposited with the American Type Culture Collection and assigned Accession Numbers PTA-296, PTA-297, and PTA-298. By “DNA sequence obtained from the overlapping clones” is intended that the DNA sequence of the catabolic gene cluster can be obtained by sequencing 18 individual clones which together comprise the entire fumonisin catabolic cluster.

Further provided are polypeptides having an amino acid sequence encoded by a nucleic acid molecule set forth in SEQ ID NO:1, the nucleic acid molecule deposited as overlapping clones with the American Type Culture Collection and assigned Accession Numbers PTA-296, PTA-297, and PTA-298, and fragments and variants thereof.

Eighteen plasmids containing overlapping clones were deposited with the American Type Culture Collection, Manassas, Va., and assigned Accession Numbers PTA-298, PTA-296, and PTA-297 (strain designations SuperCos — 8.5 — 1, BAM_PL1, and BAM_PL2, respectively). It is noted, however, that the 18 clones contain common sequences at the regions where they overlap. One of skill in the art by sequencing the clones and aligning the overlap may obtain the entire sequence of the fumonisin catabolic gene cluster. These deposits will be maintained under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure. These deposits were made merely as a convenience for those of skill in the art and are not an admission that a deposit is required under 35 U.S.C. § 112.

In addition, the present invention provides for isolated nucleic acid molecules comprising nucleotide sequences encoding the amino acid sequences of a regulatory protein (fccJ) (SEQ ID NO: 21); a carboxylesterase (fccA) (SEQ ID NO: 3), a flavin monooxygenase (fccC) (SEQ ID NO: 7); an aldehyde dehydrogenase (fccD) (SEQ ID NO: 9); an alcohol dehydrogenase (fccE) (SEQ ID NO: 11); a permease (fccI) (SEQ ID NO: 19); a CoA Ligase (fccF) (SEQ ID NO: 13); an acetohydroxyacid synthase (fccG) (SEQ ID NO: 15); a vitamin B12 transporter (fccH) (SEQ ID NO: 17); a citrate transport homolog (fccB) (SEQ ID NO: 5); a fumarate reductase (fccK) (SEQ ID NO: 23), a TonB dependent receptor (fccL) (SEQ ID NO: 25); a carbohydrate regulatory gene (fccO) (SEQ ID NO: 31); a citrate utilization B protein (fccN) (SEQ ID NO: 29); a possible flavoenzyme (fccP) (SEQ ID NO: 33); a leucine responsive regulatory protein (fccQ) (SEQ ID NO: 35); a possible N-methyl transferase (fccM) (SEQ ID NO: 27); fccR (SEQ ID NO: 37); fccS (SEQ ID NO: 39); and fccT (SEQ ID NO: 41).

Further provided are polypeptides having an amino acid sequence encoded by a nucleic acid molecule described herein, for example those set forth in SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, and 40. The nucleotide sequences of the invention can be used to isolate related sequences that are also encompassed by the present invention.

The invention encompasses isolated or substantially purified nucleic acid or protein compositions. An “isolated” or “purified” nucleic acid molecule or protein, or biologically active portion thereof, is substantially free of other cellular material, or culture medium when produced by recombinant techniques, or substantially free of chemical precursors or other chemicals when chemically synthesized. Preferably, an “isolated” nucleic acid is free of sequences (preferably protein encoding sequences) that naturally flank the nucleic acid (i.e., sequences located at the 5′ and 3′ ends of the nucleic acid) in the genomic DNA of the organism from which the nucleic acid is derived. For example, in various embodiments, the isolated nucleic acid molecule can contain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb, or 0.1 kb of nucleotide sequences that naturally flank the nucleic acid molecule in genomic DNA of the cell from which the nucleic acid is derived. A protein that is substantially free of cellular material includes preparations of protein having less than about 30%, 20%, 10%, 5%, (by dry weight) of contaminating protein. When the protein of the invention or biologically active portion thereof is recombinantly produced, preferably culture medium represents less than about 30%, 20%, 10%, or 5% (by dry weight) of chemical precursors or non-protein-of-interest chemicals.

The isolated material optionally comprises material not found with the material in its natural environment or, if the material is in its natural environment, the material has been synthetically (non-naturally) altered by deliberate human intervention to a composition and/or placed at a locus in the cell (e.g., genome or subcellular organelle) not native to a material found in that environment. The alteration to yield the synthetic material can be performed on the material within or removed from its natural state. For example, a naturally occurring nucleic acid becomes an isolated nucleic acid if it is altered, or if it is transcribed from DNA that has been altered, by non-natural, synthetic (i.e., “man-made”) methods performed within the cell from which it originates. See, e.g., Compounds and Methods for Site Directed Mutagenesis in Eukaryotic Cells, Kmiec, U.S. Pat. No. 5,565,350; In vivo Homologous Sequence Targeting in Eukaryotic Cells; Zarling et al., PCT/US93/03868. Likewise, a naturally occurring nucleic acid (e.g., a promoter) becomes isolated if it is introduced by non-naturally occurring means to a locus of the genome not native to that nucleic acid. Nucleic acids that are “isolated” as defined herein are also referred to as “heterologous” nucleic acids.

A carboxylesterase and amine oxidase from Exophiala spinifera have been previously described in U.S. Pat. No. 5,716,820 and pending U.S. patent application Ser. Nos. 08/888,949 and 08/888,950. Such disclosures are herein incorporated by reference.

The sequences of the invention can be used in combination with those previously disclosed in U.S. provisional application numbers 60/092,936 and 60/135,391 herein incorporated by reference. The enzymes and nucleotide sequences of the present invention provide a means for catabolism of fumonisin and of the fumonisin-degradation products.

The practice of the present invention will employ, unless otherwise indicated, conventional techniques of botany, microbiology, tissue culture, molecular biology, chemistry, biochemistry, and recombinant DNA technology, which are within the skill of the art. Such techniques are explained fully in the literature. See, e.g., Langenheim and Thimann (1982) Botany: Plant Biology and Its Relation to Human Affairs (John Wiley); Vasil, ed. (1984) Cell Culture and Somatic Cell Genetics of Plants, Vol. 1; Stanier et al. (1986) The Microbial World (5th ed., Prentice-Hall); Dhringra and Sinclair (1985) Basic Plant Pathology Methods (CRC Press); Maniatis et al. (1982) Molecular Cloning: A Laboratory Manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York); Glover, ed. (1985) DNA Cloning, Vols. I and II; Gait, ed. (1984) Oligonucleotide Synthesis; Hames and Higgins, eds. (1984) Nucleic Acid Hybridization; and the series Methods in Enzymology (Colowick and Kaplan, eds., Academic Press, Inc.).

In describing the present invention, the following terms will be employed, and are intended to be defined as indicated below.

By “microbe” is meant any microorganism (including both eukaryotic and prokaryotic microorganisms), such as fungi, yeast, bacteria, actinomycetes, algae, and protozoa, as well as other unicellular structures.

A “fumonisin-producing microbe” is any microbe capable of producing the mycotoxin fumonisin or analogues thereof. Such microbes are generally members of the fungal genus Fusarium, as well as recombinantly derived organisms that have been genetically altered to enable them to produce fumonisin or analogues thereof.

By “degrading or catabolizing fumonisin” is meant any modification to the fumonisin or AP1 molecule that causes a decrease or loss in its toxic activity. Such a change can comprise cleavage of any of the various bonds, oxidation, reduction, the addition or deletion of a chemical moiety, or any other change that affects the activity of the molecule. In a preferred embodiment, the modification includes hydrolysis of the two ester linkages in the molecule as a first step and then oxidative deamination. Furthermore, chemically altered fumonisin can be isolated from cultures of microbes that produce an enzyme of this invention, such as by growing the organisms in media containing radioactively-labeled fumonisin, tracing the label, and isolating the degraded toxin for further study. The degraded fumonisin can be compared to the active compound for its phytotoxicity or mammalian toxicity in known sensitive species, such as porcines and equines. Such toxicity assays are known in the art. For example, in plants a whole leaf bioassay can be used in which solutions of the active and inactive compound are applied to the leaves of sensitive plants. The leaves may be treated in situ or, alternatively, excised leaves may be used. The relative toxicity of the compounds can be estimated by grading the ensuing damage to the plant tissues and by measuring the size of lesions formed within a given time period. Other known assays can be performed at the cellular level, employing standard tissue culture methodologies e.g., using cell suspension cultures. For purposes of the invention, the fumonisin or fumonisin degradation products will be degraded to at least about 50% to about 10% or less of the original toxicity, preferably about 30% to about 5% or less, more preferably about 20% to about 1% or less.

By “fumonisin esterase” is meant any enzyme capable of hydrolysis of the ester linkages in fumonisin. Two examples of such enzymes are ESP1 and BEST1 found in U.S. Pat. No. 5,716,820 and pending U.S. application Ser. Nos. 08/888,949 and 08/888,950, both filed Jul. 7, 1997.

By “structurally related mycotoxin” is meant any mycotoxin having a chemical structure related to a fumonisin such as fumonisin B1, for example AAL toxin, fumonisin B2, fumonisin B3, fumonisin B4, fumonisin C1, fumonisin A1 and A2, and their analogues, as well as other mycotoxins having similar chemical structures that would be expected to be detoxified by activity of the fumonisin degradative enzymes elaborated by Exophiala spinifera, American Type Culture Collection Accession No. 74269, Rhinocladiella atrovirens, American Type Culture Collection Accession No. 74270, or the bacterium of American Type Culture Collection Accession No. 55552.

By “amplified” is meant the construction of multiple copies of a nucleic acid sequence or multiple copies complementary to the nucleic acid sequence using at least one of the nucleic acid sequences as a template. Amplification systems include the polymerase chain reaction (PCR) system, ligase chain reaction (LCR) system, nucleic acid sequence based amplification (NASBA, Cangene, Mississauga, Ontario), Q-Beta Replicase systems, transcription-based amplification system (TAS), and strand displacement amplification (SDA). See, e.g., Persing et al., ed. (1993) Diagnostic Molecular Microbiology: Principles and Applications, (American Society for Microbiology, Washington, D.C.). The product of amplification is termed an amplicon.

Fragments and variants of the disclosed nucleotide sequences and proteins encoded thereby are also encompassed by the present invention. By “fragment” is intended a portion of the nucleotide sequence or a portion of the amino acid sequence and hence protein encoded thereby. Fragments of a nucleotide sequence may encode protein fragments that retain the biological activity of the native protein and hence degrade or catabolize fumonisin. Alternatively, fragments of a nucleotide sequence that are useful as hybridization probes generally do not encode protein fragments retaining biological activity. Thus, fragments of a nucleotide sequence may range from at least about 20 nucleotides, about 50 nucleotides, about 100 nucleotides, and up to the full-length nucleotide sequence encoding the proteins of the invention.

A fragment of a fumonisin-degrading nucleotide sequence that encodes a biologically active portion of a fumonisin-degrading protein of the invention will encode at least 15, 20, 25, 30, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, or 950 contiguous amino acids, or up to the total number of amino acids present in a full-length fumonisin-degrading protein of the invention. By fragment it is also intended any nucleotide sequence of the fumonisin-catabolic gene cluster that encodes a biologically active portion of a fumonisin-degrading protein. Fragments of a fumonisin-degrading nucleotide sequence that are useful as hybridization probes for PCR primers generally need not encode a biologically active portion of a fumonisin-degrading protein.

Thus, a fragment of a fumonisin-degrading nucleotide sequence or a fragment of the fumonisin-catabolic gene cluster may encode a biologically active portion of a fumonisin-degrading protein or it may be a fragment that can be used as a hybridization probe or PCR primer using methods disclosed below. A biologically active portion of a fumonisin-degrading protein can be prepared by isolating a portion of one of the fumonisin-degrading nucleotide sequences or a portion of the fumonisin-catabolic gene cluster of the invention, expressing the encoded portion of the fumonisin-degrading protein (e.g., by recombinant expression in vitro), and assessing the activity of the encoded portion of the fumonisin-degrading protein. Nucleic acid molecules that are fragments of a fumonisin-degrading nucleotide sequence comprise at least 15, 20, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1,000, 1,100, 1,200, 1,300, 1,400, 1,500, 1,600, 1,700, 1,800, 1,900, 2,000, 2,100, 2,200, 2,300, 2,400, 2,500, 2,600, 2,700, 2,800 nucleotides, or up to the number of nucleotides present in a fumonisin-degrading nucleotide sequence or up to the number of nucleotides present in the full-length fumonisin catabolic gene cluster disclosed herein.

By “variants” is intended substantially similar sequences. For nucleotide sequences conservative variants include those sequences that, because of the degeneracy of the genetic code, encode the amino acid sequence of one of the fumonisin-degrading polypeptides of the invention. Naturally occurring allelic variants such as these can be identified with the use of well-known molecular biology techniques, as, for example, with polymerase chain reaction (PCR) and hybridization techniques as outlined below. Variant nucleotide sequences also include synthetically derived nucleotide sequences, such as those generated, for example, by using site-directed mutagenesis but which still encode a fumonisin-degrading protein of the invention. Generally, nucleotide sequence variants of the invention will have at least 40%, 50%, 60%, 70%, generally, 80%, preferably 85%, 90%, up to 95%, 98% sequence identity to its respective native nucleotide sequence.

By “variant” protein is intended a protein derived from the native protein by deletion (so-called truncation) or addition of one or more amino acids to the N-terminal and/or C-terminal end of the native protein; deletion or addition of one or more amino acids at one or more sites in the native protein; or substitution of one or more amino acids at one or more sites in the native protein. Such variants may result from, for example, genetic polymorphism or from human manipulation.

The proteins of the invention may be altered in various ways including amino acid substitutions, deletions, truncations, and insertions. Methods for such manipulations are generally known in the art. For example, amino acid sequence variants of the fumonisin-degrading proteins can be prepared by mutations in the DNA. Methods for mutagenesis and nucleotide sequence alterations are well known in the art. See, for example, Kunkel (1985) Proc. Natl. Acad. Sci. USA 82:488-492; Kunkel et al. (1987) Methods in Enzymol. 154:367-382; U.S. Pat. No. 4,873,192; Walker and Gaastra, eds. (1 983) Techniques in Molecular Biology (MacMillan Publishing Company, New York) and the references cited therein. Guidance as to appropriate amino acid substitutions that do not affect biological activity of the protein of interest may be found in the model of Dayhoff et al. (1978) Atlas of Protein Sequence and Structure (Natl. Biomed. Res. Found., Washington, D.C.), herein incorporated by reference. Conservative substitutions, such as exchanging one amino acid with another having similar properties, may be preferred.

Thus, the genes and nucleotide sequences of the invention include both the naturally occurring sequences as well as mutant forms. Likewise, the proteins of the invention encompass both naturally occurring proteins as well as variations and modified forms thereof. Such variants will continue to possess the desired ability to degrade fumonisin and fumonisin-like compounds. Obviously, the mutations that will be made in the DNA encoding the variant must not place the sequence out of the reading frame and preferably will not create complementary regions that could produce secondary mRNA structure. See, EP Patent Application Publication No. 75,444.

The deletions, insertions, and substitutions of the protein sequences encompassed herein are not expected to produce radical changes in the characteristics of the protein. However, when it is difficult to predict the exact effect of the substitution, deletion, or insertion in advance of doing so, one skilled in the art will appreciate that the effect will be evaluated by routine screening assays. That is, the activity can be evaluated by a decrease or loss in the toxic activity of fumonisin or AP1.

Variant nucleotide sequences and proteins also encompass sequences and proteins derived from a mutagenic and recombinogenic procedure such as DNA shuffling. With such a procedure, one or more different fumonisin-degrading protein coding sequences can be manipulated to create a new fumonisin-degrading protein possessing the desired properties. In this manner, libraries of recombinant polynucleotides are generated from a population of related sequence polynucleotides comprising sequence regions that have substantial sequence identity and can be homologously recombined in vitro or in vivo. For example, using this approach, sequence motifs encoding a domain of interest may be shuffled between the fumonisin-degrading genes of the invention and other known fumonisin-degrading genes to obtain a new gene coding for a protein with an improved property of interest, such as an increased K m in the case of an enzyme. Strategies for such DNA shuffling are known in the art. See, for example, Stemmer (1994) Proc. Natl. Acad. Sci. USA 91:10747-10751; Stemmer (1994) Nature 370:389-391; Crameri et al. (1997) Nature Biotech. 15:436-438; Moore et al. (1997) J. Mol. Biol. 272:336-347; Zhang et al. (1997) Proc. Natl. Acad. Sci. USA 94:4504-4509; Crameri et al. (1998) Nature 391:288-291; and U.S. Pat. Nos. 5,605,793 and 5,837,458.

When the nucleic acid is prepared or altered synthetically, advantage can be taken of known codon preferences of the intended host where the nucleic acid is to be expressed. For example, although nucleic acid sequences of the present invention may be expressed in both monocotyledonous and dicotyledonous plant species, sequences can be modified to account for the specific codon preferences and GC content preferences of monocotyledonous plants or dicotyledonous plants as these preferences have been shown to differ (Murray et al. (1989) Nucl. Acids Res. 17:477-498). Thus, the maize preferred codon for a particular amino acid may be derived from known gene sequences from maize. Maize codon usage for 28 genes from maize plants are listed in Table 4 of Murray et al., supra.

As used herein, “heterologous” in reference to a nucleic acid is a nucleic acid that originates from a foreign species, or, if from the same species, is substantially modified from its native form in composition and/or genomic locus by deliberate human intervention. For example, a promoter operably linked to a heterologous structural gene is from a species different from that from which the structural gene was derived, or, if from the same species, one or both are substantially modified from their original form. A heterologous protein may originate from a foreign species or, if from the same species, is substantially modified from its original form by deliberate human intervention.

By “host cell” is meant a cell that contains a vector and supports the replication and/or expression of the expression vector. Host cells may be prokaryotic cells such as E. coli, or eukaryotic cells such as yeast, insect, amphibian, or mammalian cells. Preferably, host cells are monocotyledonous or dicotyledonous plant cells, including but not limited to maize, sorghum, sunflower, soybean, wheat, alfalfa, rice, cotton, and tomato. A particularly preferred monocotyledonous host cell is a maize host cell.

The term “hybridization complex” includes reference to a duplex nucleic acid structure formed by two single-stranded nucleic acid sequences selectively hybridized with each other.

As used herein, “operably linked” includes reference to a functional linkage between a promoter and a second sequence, wherein the promoter sequence initiates and mediates transcription of the DNA sequence corresponding to the second sequence. Generally, operably linked means that the nucleic acid sequences being linked are contiguous and, where necessary to join two protein coding regions, contiguous and in the same reading frame.

As used herein, “polynucleotide” includes reference to a deoxyribopolynucleotide, ribopolynucleotide, or analogues thereof that have the essential nature of a natural ribonucleotide in that they hybridize, under stringent hybridization conditions, to substantially the same nucleotide sequence as naturally occurring nucleotides and/or allow translation into the same amino acid(s) as the naturally occurring nucleotide(s). A polynucleotide can be full-length or a subsequence of a native or heterologous structural or regulatory gene. Unless otherwise indicated, the term includes reference to the specified sequence as well as the complementary sequence thereof. Thus, DNAs or RNAs with backbones modified for stability or for other reasons are “polynucleotides” as that term is intended herein. Moreover, DNAs or RNAs comprising unusual bases, such as inosine, or modified bases, such as tritylated bases, to name just two examples, are polynucleotides as the term is used herein. It will be appreciated that a great variety of modifications have been made to DNA and RNA that serve many useful purposes known to those of skill in the art. The term polynucleotide as it is employed herein embraces such chemically, enzymatically, or metabolically modified forms of polynucleotides, as well as the chemical forms of DNA and RNA characteristic of viruses and cells, including inter alia, simple and complex cells.

As used herein, “promoter” includes reference to a region of DNA upstream from the start of transcription and involved in recognition and binding of RNA polymerase and other proteins to initiate transcription. A “plant promoter” is a promoter capable of initiating transcription in plant cells. Exemplary plant promoters include, but are not limited to, those that are obtained from plants, plant viruses, and bacteria that comprise genes expressed in plant cells, such as Agrobacterium or Rhizobium. Examples are promoters that preferentially initiate transcription in certain tissues, such as leaves, roots, seeds, fibers, xylem vessels, tracheids, or sclerenchyma. Such promoters are referred to as “tissue preferred”. A “cell type” specific promoter primarily drives expression in certain cell types in one or more organs, for example, vascular cells in roots or leaves. An “inducible” promoter is a promoter that is under environmental control. Examples of environmental conditions that may effect transcription by inducible promoters include anaerobic conditions or the presence of light. Another type of promoter is a developmentally regulated promoter. For example, a promoter that drives expression during pollen development. Tissue-preferred, cell type specific, developmentally regulated, and inducible promoters constitute the class of “non-constitutive” promoters. A “constitutive” promoter is a promoter that is active under most environmental conditions. Constitutive promoters are known in the art and include, for example, 35S promoter (Meyer et al. (1997) J. Gen. Virol. 78:3147-3151); ubiquitin; as well as those disclosed in U.S. Pat. Nos. 5,608,149; 5,608,144; 5,604,121; 5,569,597; 5,466,785; 5,399,680; 5,268,463; 5,608,142.

As used herein, “recombinant” includes reference to a cell or vector that has been modified by the introduction of a heterologous nucleic acid or that the cell is derived from a cell so modified. Thus, for example, recombinant cells express genes that are not found in identical form within the native (non-recombinant) form of the cell or express native genes that are otherwise abnormally expressed, underexpressed, or not expressed at all as a result of deliberate human intervention. The term “recombinant” as used herein does not encompass the alteration of the cell or vector by naturally occurring events (e.g., spontaneous mutation, natural transformation/transduction/transposition) such as those occurring without deliberate human intervention.

As used herein, a “recombinant expression cassette” is a nucleic acid construct, generated recombinantly or synthetically, with a series of specified nucleic acid elements that permit transcription of a particular nucleic acid in a target cell. The recombinant expression cassette can be incorporated into a plasmid, chromosome, mitochondrial DNA, plastid DNA, virus, or nucleic acid fragment. Typically, the recombinant expression cassette portion of an expression vector includes, among other sequences, a nucleic acid to be transcribed, and a promoter.

The term “selectively hybridizes” includes reference to hybridization, under stringent hybridization conditions, of a nucleic acid sequence to a specified nucleic acid target sequence to a detectably greater degree (e.g., at least 2-fold over background) than its hybridization to non-target nucleic acid sequences and to the substantial exclusion of non-target nucleic acids. Selectively hybridizing sequences typically have about at least 40 to 80% sequence identity, preferably 90% sequence identity, and most preferably 100% sequence identity (i.e., complementary) with each other.

The nucleotide sequences of the invention can be used to isolate corresponding sequences from other organisms. In this manner, methods such as PCR, hybridization, and the like can be used to identify such sequences based on their sequence homology to the sequences set forth herein. Sequences isolated based on their sequence identity to the entire fumonisin-degrading coding sequences or to the catabolic gene cluster sequence set forth herein or to fragments thereof are encompassed by the present invention.

In a PCR approach, oligonucleotide primers can be designed for use in PCR reactions to amplify corresponding DNA sequences from cDNA or genomic DNA extracted from any organism of interest. Methods for designing PCR primers and PCR cloning are generally known in the art and are disclosed in Sambrook et al. (1989) Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, Plainview, N.Y.). See also Innis et al., eds. (1990) PCR Protocols: A Guide to Methods and Applications (Academic Press, New York); Innis and Gelfand, eds. (1995) PCR Strategies (Academic Press, New York); and Innis and Gelfand, eds. (1999) PCR Methods Manual (Academic Press, New York). Known methods of PCR include, but are not limited to, methods using paired primers, nested primers, single specific primers, degenerate primers, gene-specific primers, vector-specific primers, partially-mismatched primers, and the like.

In hybridization techniques, all or part of a known nucleotide sequence is used as a probe that selectively hybridizes to other corresponding nucleotide sequences present in a population of cloned genomic DNA fragments or cDNA fragments (i.e., genomic or cDNA libraries) from a chosen organism. The hybridization probes may be genomic DNA fragments, cDNA fragments, RNA fragments, or other oligonucleotides, and may be labeled with a detectable group such as 32 P, or any other detectable marker. Thus, for example, probes for hybridization can be made by labeling synthetic oligonucleotides based on the fumonisin-degrading coding sequences of the invention. Methods for preparation of probes for hybridization and for construction of cDNA and genomic libraries are generally known in the art and are disclosed in Sambrook et al. (1989) Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, Plainview, N.Y.).

For example, an entire fumonisin-degrading nucleotide sequence or the entire fumonisin catabolic gene cluster sequence disclosed herein, or one or more portions thereof, may be used as a probe capable of specifically hybridizing to corresponding fumonisin-degrading sequences and messenger RNAs. To achieve specific hybridization under a variety of conditions, such probes include sequences that are unique among fumonisin-degrading sequences and are preferably at least about 10 nucleotides in length, and most preferably at least about 20 nucleotides in length. Such probes may be used to amplify corresponding fumonisin-degrading sequences from a chosen organism by PCR. This technique may be used to isolate additional coding sequences from a desired plant organism or as a diagnostic assay to determine the presence of coding sequences in a plant an organism. Hybridization techniques include hybridization screening of plated DNA libraries (either plaques or colonies; see, for example, Sambrook et al. (1989) Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, Plainview, N.Y.).

Hybridization of such sequences may be carried out under stringent conditions. By “stringent conditions” or “stringent hybridization conditions” is intended conditions under which a probe will hybridize to its target sequence to a detectably greater degree than to other sequences (e.g., at least 2-fold over background). Stringent conditions are sequence-dependent and will be different in different circumstances. By controlling the stringency of the hybridization and/or washing conditions, target sequences that are 100% complementary to the probe can be identified (homologous probing). Alternatively, stringency conditions can be adjusted to allow some mismatching in sequences so that lower degrees of similarity are detected (heterologous probing). Generally, a probe is less than about 1000 nucleotides in length, preferably less than 500 nucleotides in length.

Typically, stringent conditions will be those in which the salt concentration is less than about 1.5 M Na ion, typically about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30° C. for short probes (e.g., 10 to 50 nucleotides) and at least about 60° C. for long probes (e.g., greater than 50 nucleotides). Stringent conditions may also be achieved with the addition of destabilizing agents such as formamide. Exemplary low stringency conditions include hybridization with a buffer solution of 30 to 35% formamide, 1 M NaCl, 1% SDS (sodium dodecyl sulphate) at 37° C., and a wash in 1× to 2× SSC (20× SSC=3.0 M NaCl/0.3 M trisodium citrate) at 50 to 55° C. Exemplary moderate stringency conditions include hybridization in 40 to 45% formamide, 1.0 M NaCl, 1% SDS at 37° C., and a wash in 0.5× to 1× SSC at 55 to 60° C. Exemplary high stringency conditions include hybridization in 50% formamide, 1 M NaCl, 1% SDS at 37° C., and a wash in 0.1× SSC at 60 to 65° C.

Specificity is typically the function of post-hybridization washes, the critical factors being the ionic strength and temperature of the final wash solution. For DNA—DNA hybrids, the T m can be approximated from the equation of Meinkoth and Wahl (1984) Anal. Biochem. 138:267-284: T m =81.5° C.+16.6 (log M)+0.41 (% GC)−0.61 (% form)−500/L; where M is the molarity of monovalent cations, % GC is the percentage of guanosine and cytosine nucleotides in the DNA, % form is the percentage of formamide in the hybridization solution, and L is the length of the hybrid in base pairs. The T m is the temperature (under defined ionic strength and pH) at which 50% of a complementary target sequence hybridizes to a perfectly matched probe. T m is reduced by about 1° C. for each 1% of mismatching; thus, T m , hybridization, and/or wash conditions can be adjusted to hybridize to sequences of the desired identity. For example, if sequences with ≧90% identity are sought, the T m can be decreased 10° C. Generally, stringent conditions are selected to be about 5° C. lower than the thermal melting point (T m ) for the specific sequence and its complement at a defined ionic strength and pH. However, severely stringent conditions can utilize a hybridization and/or wash at 1, 2, 3, or 4° C. lower than the thermal melting point (T m ); moderately stringent conditions can utilize a hybridization and/or wash at 6, 7, 8, 9, or 10° C. lower than the thermal melting point (T m ); low stringency conditions can utilize a hybridization and/or wash at 11, 12, 13, 14, 15, or 20° C. lower than the thermal melting point (T m ). Using the equation, hybridization and wash compositions, and desired T m , those of ordinary skill will understand that variations in the stringency of hybridization and/or wash solutions are inherently described. If the desired degree of mismatching results in a T m of less than 45° C. (aqueous solution) or 32° C. (formamide solution), it is preferred to increase the SSC concentration so that a higher temperature can be used. An extensive guide to the hybridization of nucleic acids is found in Tijssen (1993) Laboratory Techniques in Biochemistry and Molecular Biology—Hybridization with Nucleic Acid Probes, Part I, Chapter 2 (Elsevier, N.Y.); and Ausubel et al., eds. (1995) Current Protocols in Molecular Biology, Chapter 2 (Greene Publishing and Wiley-Interscience, New York). See Sambrook et al. (1989) Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, Plainview, N.Y.).

In general, sequences that encode for a fumonisin-degrading protein and hybridize to the fumonisin-degrading sequences or the sequences of the fumonisin catabolic gene cluster disclosed herein will be at least 40% to 50% homologous, about 60% to 70% homologous, and even about 80%, 85%, 90%, 95% to 98% homologous or more with the disclosed sequences. That is, the sequence similarity of sequences may range, sharing at least about 40% to 50%, about 60% to 70%, and even about 80%, 85%, 90%, 95% to 98% sequence similarity.

The following terms are used to describe the sequence relationships between two or more nucleic acids or polynucleotides: (a) “reference sequence”, (b) “comparison window”, (c) “sequence identity”, (d) “percentage of sequence identity”, and (e) “substantial identity”.

(a) As used herein, “reference sequence” is a defined sequence used as a basis for sequence comparison. A reference sequence may be a subset or the entirety of a specified sequence; for example, as a segment of a full-length cDNA or gene sequence, or the complete cDNA or gene sequence.

(b) As used herein, “comparison window” makes reference to a contiguous and specified segment of a polynucleotide sequence, wherein the polynucleotide sequence in the comparison window may comprise additions or deletions (i.e., gaps) compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two sequences. Generally, the comparison window is at least 20 contiguous nucleotides in length, and optionally can be 30, 40, 50, 100, or longer. Those of skill in the art understand that to avoid a high similarity to a reference sequence due to inclusion of gaps in the polynucleotide sequence a gap penalty is typically introduced and is subtracted from the number of matches.

Methods of alignment of sequences for comparison are well known in the art. Optimal alignment of sequences for comparison may be conducted by the local homology algorithm of Smith et al. (1981) Adv. Appl. Math. 2:482; by the homology alignment algorithm of Needleman et al. ( 1970) J. Mol. Biol. 48:443; by the search for similarity method of Pearson et al. (1988) Proc. Natl. Acad. Sci. 85:2444; by computerized implementations of these algorithms, including, but not limited to: CLUSTAL in the PC/Gene program by Intelligenetics, Mountain View, Calif.; GAP, BESTFIT, BLAST, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group (GCG), 575 Science Drive, Madison, Wis., USA; the CLUSTAL program is well described by Higgins et al. (1988) Gene 73:237-244 (1988); Higgins et al. (1989) CABIOS 5:151-153; Corpet et al. (1988) Nucleic Acids Res. 16:10881-90; Huang et al. (1992) Computer Applications in the Biosciences 8:155-65, and Person et al. (1994) Meth. Mol. Biol. 24:307-331; preferred computer alignment methods also include the BLASTP, BLASTN, and BLASTX algorithms (see Altschul et al. (1990) J. Mol. Biol. 215:403-410). Alignment is also often performed by inspection and manual alignment.

(c) As used herein, “sequence identity” or “identity” in the context of two nucleic acid or polypeptide sequences makes reference to the residues in the two sequences that are the same when aligned for maximum correspondence over a specified comparison window. When percentage of sequence identity is used in reference to proteins it is recognized that residue positions which are not identical often differ by conservative amino acid substitutions, where amino acid residues are substituted for other amino acid residues with similar chemical properties (e.g., charge or hydrophobicity) and therefore do not change the functional properties of the molecule. When sequences differ in conservative substitutions, the percent sequence identity may be adjusted upwards to correct for the conservative nature of the substitution. Sequences that differ by such conservative substitutions are said to have “sequence similarity” or “similarity”. Means for making this adjustment are well known to those of skill in the art. Typically this involves scoring a conservative substitution as a partial rather than a full mismatch, thereby increasing the percentage sequence identity. Thus, for example, where an identical amino acid is given a score of 1 and a non-conservative substitution is given a score of zero, a conservative substitution is given a score between zero and 1. The scoring of conservative substitutions is calculated, e.g., as implemented in the program PC/GENE (Intelligenetics, Mountain View, Calif.).

(d) As used herein, “percentage of sequence identity” means the value determined by comparing two optimally aligned sequences over a comparison window, wherein the portion of the polynucleotide sequence in the comparison window may comprise additions or deletions (i.e., gaps) as compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two sequences. The percentage is calculated by determining the number of positions at which the identical nucleic acid base or amino acid residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison, and multiplying the result by 100 to yield the percentage of sequence identity.

(e)(i) The term “substantial identity” of polynucleotide sequences means that a polynucleotide comprises a sequence that has at least 70% sequence identity, preferably at least 80%, more preferably at least 90%, and most preferably at least 95%, compared to a reference sequence using one of the alignment programs described using standard parameters. One of skill in the art will recognize that these values can be appropriately adjusted to determine corresponding identity of proteins encoded by two nucleotide sequences by taking into account codon degeneracy, amino acid similarity, reading frame positioning, and the like. Substantial identity of amino acid sequences for these purposes normally means sequence identity of at least 60%, more preferably at least 70%, 80%, 90%, and most preferably at least 95%.

Another indication that nucleotide sequences are substantially identical is if two molecules hybridize to each other under stringent conditions. Generally, stringent conditions are selected to be about 5° C. lower than the thermal melting point (T m ) for the specific sequence at a defined ionic strength and pH. However, stringent conditions encompass temperatures in the range of about 1° C. to about 20° C., depending upon the desired degree of stringency as otherwise qualified herein. Nucleic acids that do not hybridize to each other under stringent conditions are still substantially identical if the polypeptides they encode are substantially identical. This may occur, e.g., when a copy of a nucleic acid is created using the maximum codon degeneracy permitted by the genetic code. One indication that two nucleic acid sequences are substantially identical is when the polypeptide encoded by the first nucleic acid is immunologically cross reactive with the polypeptide encoded by the second nucleic acid.

(e)(ii) The term “substantial identity” in the context of a peptide indicates that a peptide comprises a sequence with at least 70% sequence identity to a reference sequence, preferably 80%, more preferably 85%, most preferably at least 90% or 95% sequence identity to the reference sequence over a specified comparison window. Preferably, optimal alignment is conducted using the homology alignment algorithm of Needleman et al. (1970) J. Mol. Biol. 48:443. An indication that two peptide sequences are substantially identical is that one peptide is immunologically reactive with antibodies raised against the second peptide. Thus, a peptide is substantially identical to a second peptide, for example, where the two peptides differ only by a conservative substitution. Peptides that are “substantially similar” share sequences as noted above except that residue positions that are not identical may differ by conservative amino acid changes.

As indicated, the enzymes and nucleotide sequences encoding such enzymes are involved in the degradation of fumonisin and fumonisin-like compounds. Such enzymes and nucleotide sequences can be utilized alone or in combination to engineer microbes or other organisms to completely metabolize fumonisin and resist its toxic effects. That is, at least one of the genes of the invention can be used to transform a host cell of interest. Alternatively, the entire fumonisin catabolic cluster can be cloned into the host cell. Methods for cloning gene clusters into a plasmid, a cosmid or a Bac are known in the art. The cloned cluster can then be moved to the host cell of interest, generally another microorganism or yeast cell. Under suitable induction conditions, the pathway will be activated and fumonisin degraded. See, for example, Prieto et al. (1996) J. Bacteriol. 178(1):111-120, herein incorporated by reference.

Fumonisin is produced in the intercellular spaces (apoplast) of Fusarium-infected maize cells. Thus, the apoplast is the preferred location for esterase and possibly other catabolic enzymes. It is possible that some fumonisin could diffuse or be transported into the maize cells before it is broken down by the apoplastic enzymes and may escape catabolism. Thus, it may be beneficial to express a fumonisin pump in such cells. In this manner, any fumonisin entering the cell will be pumped out and reexposed to catabolic enzymes. Similar toxin pumps exist in other toxin-producing fungi that show resistance to toxins or antibiotics.

The enzymes from Bacterium 2412.1 involved in the degradation of fumonisin are provided in Table 1. Additionally, they are discussed briefly below.

TABLE 1
			Predicted ORF
Gene	Nucleotides	Frame	Size	BLASTP Homology	Functional Class
fccA	13803 to 15389	+3	529 aa	Esterase	Metabolism
fccB	15627 to 16913	+3	429 aa	Citrate Transport	Transport
fccC	19074 to 20747	+3	558 aa	Flavin monoox	Metabolism
fccD	11698 to 10295	−3	468 aa	Aldehyde dehydrogenase	Metabolism
fccE	6435 to 7370	+3	312 aa	Alcohol dehydrogenase	Metabolism
fccF	7814 to 9187	+2	458 aa	Co-A Ligase	Metabolism
fccG	13474 to 11774	−2	567 aa	Acetohydroxy acid synthase	Metabolism
fccH	24222 to 21361	−3	954 aa	B12 Transport	Transport
fccI	17189 to 18487	−2	433 aa	Permease	Transport
fccJ	9317 to 10225	+2	303 aa	Cat Transcription Factor	Regulatory
fccK	3723 to 5111	+3	463 aa	Aspartae Oxidase (Fumarate	Metabolism
				Reductase)
fccL	979 to 3381	+1	801 aa	TonB Dependent Receptor	Transport
fccM	15557 to 14394	−1	391 aa	N-methyl transferase	Metabolism
fccN	5104 to 6282	+1	393 aa	Cit-B homolog	Metabolism
fccO	10892 to 11368	+2	159 aa	Car 2 comp Regulatory Protein	Regulatory
fccE	3408 to 2353	−3	352 aa	Possible Flavoenzyme	Metabolism
fccQ	229 to 705	+1	159 aa	LRP Regulatory Protein	Regulatory
fccR	8926 to 7854	−2	358 aa	Unknown	?
fccS	2221 to 1660	−1	183 aa	Unknown	?
fccT	1590 to 1060	−1	177 aa	Unknown	?

fccA Esterase:

This esterase, also known as BacEst, was previously shown to hydrolyze the tricarballylate esters of fumonisins. This enzyme is the bacterial homolog of the Exophiala spinifera esterase, ESP1.

fccB Citrate/Tricarballylate Transport Protein:

This protein may be necessary to break down the tricarballylic acid resulting from hydrolysis of fumonisin by the esterase. The citrate/tricarballylate transporter can be used to transport the tricarballylic acid into the cell where it may be able to be broken down by endogenous enzymes. Additional enzymes that may be discovered could be used as well to effect the removal of tricarballylate once it is in the cell.

fccC Flavin Monooxygenase:

This enzyme has homology to monooxygnease that act on keto or quinone groups, oxidizing them to a carboxylate with carbon chain breakage. In our detoxification strategy, it may be necessary to provide more complete catabolism of fumonisin in transgenic maize or other transgenic organisms than can be provided by esterase and deaminase enzymes. If so, the monooxygenase activity of this clone is predicted to result in the oxidation of 2-OP to a compound that would lack a keto group, having instead a terminal aldehyde group, or possibly a carboxylate group. This is due to a type of enzymatic oxidation referred to as Baeyer-Villiger oxidation, in which monooxygen is inserted adjacent to a keto function, resulting in a lactone or ester linkage (Walsch & Chen, Angew. Chem. Int Ed. Engl 27 (1988) 333-343). The metabolism of trans-cyclohexane-1,2 diol by Acinetobacter provides a model for the activity of a Baeyer-Villiger monooxygenase on 2-OP (Davey & Trudgill, 1977, Eur. J. Biochem 74:115-127). This diol is first oxidized to ortho hydroxy cyclohexanone by a different enzyme, and then a monooxygen is inserted between the quione and hydroxy functions by the Baeyer-Villiger enzyme, cyclohexanone monooxygenase. This intermediate spontaneously rearranges to a linear aldehyde carboxylic acid. By analogy, for 2-OP we would predict insertion of oxygen between carbons 2 and 3 and then spontaneous cleavage to a C22 aldehyde and acetic acid. Further oxidation by an aldehyde dehydrogenase would convert this compound to a carboxylic acid; other transformations would also be possible given the high reactivity of the aldehyde group.

fccD Aldehyde Dehydrogenase:

One likely product of flavin monooxygenase oxidation of 2-OP is a C-20 aldehyde (see above); if so, we would predict that an aldehyde dehyrogenase would be available to oxidize the aldehyde to a carboxylate group, as a prerequisite to insertion into the beta-oxidation pathway for catabolism to acetyl co-A.

fccE Alcohol Dehydrogenase:

This gene shows homology to members of the short-chain dehydrogenase (SD) class of oxidoreductases, designated in Prosite as group PSO-00060. This class contains many NADP or NAD+ requiring alcohol dehydrogenases as well as anabolic enzymes that convert 2-keto Co-A acid esters to -corresponding alcohols. This enzyme may be responsible for oxidation of the C-5, alcohol of the carboxylate to a keto group, which would result in an alpha keto acid that could be further degraded by beta-oxidation. The enzyme could also oxidize hydroxyls at C13 and C14 to ketones, allowing a monooxygenase such as that described above to further oxidize and fragment the carbon chain. It is also possible that this enzyme attacks the C5 hydroxyl of AP1. Since the C-5 hydroxyl group of AP1 is considered important for toxicity, this enzyme may be useful in detoxification of AP1.

fccF CoA Ligase:

This enzyme may provide conjugation of a fumonisin-derived carboxylic acid with coenzyme A, prior to beta-oxidation.

fccG Acetohydroxyacid Synthase:

This gene may code for a protein that provides a pH stat when fumonisin metabolites are being metabolized.

fccH Vitamin B12 Receptor:

This receptor possibly plays a role in membrane transport of breakdown products or import of cofactors needed for enzymes in the pathway.

fccI Permease:

This protein is expected to have a role in the transport of metabolites into the cells, or exclusion of fumonisin.

fccJ Regulatory Protein:

Similar to LysR regulatory protein from E. coli , this protein probably functions to suppress transcription of pathway genes until the appropriate stimulus (i.e., fumonisin or AP1 or TCA) is present.

fccK Fumarte Reductase (Aspartate Oxidase):

This gene encodes a redox enzyme homolog that may oxidize the C-2 amine of AP1 and/or fumonisin.

fccL TonB Dependent Receptor:

This protein is involved in iron uptake.

FccM:

This protein has homology to an N-methyl transferase and may be involved in antibiotic resistance. In addition, it may also function in methylating FB1 or AP1 to reduce toxicity.

FccN:

The fccN gene encodes a citrate utilization B protein.

FccO Carbohydrate Regulatory Protein:

This protein may be involved in the regulation of carbohydrate pathways. It may also down-regulate genes for non-fumonisin catabolysis or may be involved in regulating gene expression in the fumonisin cluster.

FccP:

This gene is contained within the antisense strand of fccl and may encode a flavoenzyme. The protein may be an AP1/FB1 deaminating enzyme based on similarity to flavoenzymes which include amine oxidases.

FccQ Leucine Responsive Regulatory Protein:

This protein may be involved in regulation of catabolic gene expression.

FccR, FccS, FccT:

These proteins show no significant homology to other known proteins.

Compositions of the invention include the native nucleotide sequences for the catabolic genes, as well as variants and modifications thereof. It is recognized that having elucidated the pathway for fumonisin catabolism in Bacterium 2412.1, such DNA sequences can be inserted into expression cassettes and used to transform a variety of organisms. Enzymes produced recombinantly may be tested for their ability to modify fumonisin, fumonisin-related toxins, or a fumonisin byproduct using labeled starting material and appropriate buffer and cofactor conditions. For example, to test aldehyde dehydrogenase activity, the aldehyde dehydrogenase produced in a recombinant manner would be incubated with cofactors, NAD+ or NADP+, and 14 C-labeled 2-OP for various times and then an aliquot of the reaction mix spotted on TLC. Enzyme activity would be indicated by the appearance of a new radiolabeled spot at a different Rf on the TLC plate. Compositions of the invention include the native nucleotide sequences for the catabolic genes and the fumonisin gene cluster, as well as variants and modifications thereof. Particularly, the nucleotide sequences, or fragments thereof, of SEQ ID NOS: 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38 and 40 can be utilized to isolate and/or synthesize full-length DNA sequences. Methods are known in the art for cloning of full-length cDNAs. Such techniques include, for example, RACE-PCR using oligonucleotide primers based on the known sequence, or cDNA library screening with labeled cDNA as probe, or by means of PCR of genomic DNA using a gene-specific probe and end-ligated primers (e.g., Genome Walker™ Clontech). It is recognized that having elucidated the pathway for fumonisin catabolism in Exophiala, such DNA sequences can be inserted into expression cassettes and used to transform a variety of organisms. Enzymes produced recombinantly may be tested for their ability to modify fumonisin or a fumonisin byproduct using labeled starting material and appropriate buffer and cofactor conditions. For example, to test aldehyde dehydrogenase activity, the aldehyde dehydrogenase produced in a recombinant manner would be incubated with cofactors, NAD+ or NADP, and 14 C -labeled 2-OP for various times and then an aliquot of the reaction mix spotted on TLC. Enzyme activity would be indicated by the appearance of a new radiolabeled spot at a different Rf on the TLC plate.

The sequences of the invention can be introduced into any host organism. The sequences to be introduced may be used in expression cassettes for expression in the host of interest where expression in the host is necessary for transcription.

One of skill would recognize that modifications can be made to a protein of the present invention without diminishing its biological activity. Some modifications may be made to facilitate the cloning, expression, or incorporation of the targeting molecule into a fusion protein. Such modifications are well known to those of skill in the art and include, for example, a methionine added at the amino terminus to provide an initiation site, or additional amino acids (e.g., poly His) placed on either terminus to create conveniently located restriction sites or termination codons or purification sequences.

Where expression cassettes are needed, such expression cassettes will comprise a transcriptional initiation region linked to the coding sequence or antisense sequence of the nucleotide of interest. Such an expression cassette is provided with a plurality of restriction sites for insertion of the sequence to be under the transcriptional regulation of the regulatory regions. The expression cassette may additionally contain selectable marker genes.

The marker gene confers a selectable phenotype on the transformed cells. Usually, the selectable marker gene will encode antibiotic resistance, with suitable genes including genes coding for resistance to the antibiotic spectinomycin (e.g., the aada gene), the streptomycin phosphotransferase (SPT) gene coding for streptomycin resistance, the neomycin phosphotransferase (NPTII) gene encoding kanamycin or geneticin resistance; the hygromycin phosphotransferase (HPT) gene coding for hygromycin resistance, genes coding for resistance to herbicides which act to inhibit the action of acetolactate synthase (ALS), in particular the sulfonylurea-type herbicides (e.g, the acetolactate synthase (ALS) gene containing mutations leading to such resistance in particular the S4 and/or Hra mutations), genes coding for resistance to herbicides which act to inhibit action of glutamine synthase, such as phosphinothricin or basta (e.g., the bar gene), or other such genes known in the art. The bar gene encodes resistance to the herbicide basta, and the ALS gene encodes resistance to the herbicide chlorsulfuron.

The transcriptional initiation region, the promoter, may be native or analogous or foreign or heterologous to the host as well as to the coding sequence. Additionally, the promoter may be the natural sequence or alternatively a synthetic sequence. By foreign it is intended that the transcriptional initiation region is not found in the native plant into which the transcriptional initiation region is introduced. As used herein a chimeric gene comprises a coding sequence operably linked to a transcription initiation region that is heterologous to the coding sequence.

The transcriptional cassette will include in the 5′-to-3′ direction of transcription, a transcriptional and translational initiation region, a DNA sequence of interest, and a transcriptional and translational termination region functional in the host. The termination region may be native with the transcriptional initiation region, may be native with the DNA sequence of interest, or may be derived from another source. For use in plants or plant cells, convenient termination regions are available from the Ti-plasmid of A. tumefaciens, such as the octopine synthase and nopaline synthase termination regions. See also Guerineau et al. (1991) Mol. Gen. Genet. 262:141-144; Proudfoot (1991) Cell 64:671-674; Sanfacon et al. (1991) Genes Dev. 5:141-149; Mogen et al. (1990) Plant Cell. 2:1261-1272; Munroe et al. (1990) Gene 91:151-158; Ballas et al. (1989) Nucleic Acids Res. 17:7891-7903; Joshi et al. (1987) Nucleic Acids Res. 15:9627-9639.

Nucleotide sequences of the invention are provided in expression cassettes for expression in the host cell of interest. The cassette will include 5′ and 3′ regulatory sequences operably linked to the sequence of interest. The cassette may additionally contain at least one additional sequence to be cotransformed into the organism. Alternatively, the additional sequence(s) can be provided on another expression cassette.

Where appropriate, the gene(s) may be optimized for increased expression in the transformed plant. That is, the genes can be synthesized using plant-preferred codons for improved expression. Methods are available in the art for synthesizing plant-preferred genes. See, for example, U.S. Pat. Nos. 5,380,831, 5,436,391, and Murray et al. (1989) Nucleic Acids Res. 17:477-498, herein incorporated by reference.

Additional sequence modifications are known to enhance gene expression in a cellular host. These include elimination of sequences encoding spurious polyadenylation signals, exon-intron splice site signals, transposon-like repeats, and other such well-characterized sequences that may be deleterious to gene expression. The G-C content of the sequence may be adjusted to levels average for a given cellular host, as calculated by reference to known genes expressed in the host cell. When possible, the sequence is modified to avoid predicted hairpin secondary mRNA structures.

The expression cassettes may additionally contain 5′ leader sequences in the expression cassette construct. Such leader sequences can act to enhance translation. Translation leaders are known in the art and include: picornavirus leaders, for example, EMCV leader (Encephalomyocarditis 5′ noncoding region) (Elroy-Stein et al. (1989) PNAS USA 86:6126-6130); potyvirus leaders, for example, TEV leader (Tobacco Etch Virus) (Allison et al. (1986); MDMV leader (Maize Dwarf Mosaic Virus); Virology 154:9-20), and human immunoglobulin heavy-chain binding protein (BiP), (Macejak et al. (1991) Nature 353:90-94); untranslated leader from the coat protein mRNA of alfalfa mosaic virus (AMV RNA 4) (Jobling et al. (1987) Nature 325:622-625); tobacco mosaic virus leader (TMV) (Gallie et al. (1989) in Molecular Biology of RNA, ed. Cech (Liss, New York), pp. 237-256); and maize chlorotic mottle virus leader (MCMV) (Lommel et al. (1991) Virology 81:382-385). See also, Della-Cioppa et al. (1987) Plant Physiol. 84:965-968. Other methods known to enhance translation can also be utilized, for example, introns, and the like.

In preparing the expression cassette, the various DNA fragments may be manipulated, so as to provide for the DNA sequences in the proper orientation and, as appropriate, in the proper reading frame. Toward this end, adapters or linkers may be employed to join the DNA fragments or other manipulations may be involved to provide for convenient restriction sites, removal of superfluous DNA, removal of restriction sites, or the like. For this purpose, in vitro mutagenesis, primer repair, restriction, annealing, resubstitutions, e.g., transitions and transversions, may be involved.

In the same manner, a plant can be transformed with the nucleotide sequences of the invention to provide complete detoxification of fumonisin in the transformed plant and plant products. Such plants include, for example, species from the genera Cucurbita, Rosa, Vitis, Juglans, Fragaria, Lotus, Medicago, Onobrychis, Trifolium, Trigonella, Vigna, Citrus, Linum, Geranium, Manihot, Daucus, Arabidopsis, Brassica, Raphanus, Sinapis, Atropa, Capsicum, Datura, Hyoscyamus, Lycopersicon, Nicotiana, Solanum, Petunia, Digitalis, Majorana, Ciahorium, Helianthus, Lactuca, Bromus, Asparagus, Antirrhinum, Heterocallis, Nemesis, Pelargonium, Panieum, Pennisetum, Ranunculus, Senecio, Salpiglossis, Cucumis, Browaalia, Glycine, Pisum, Phaseolus, Lolium, Oryza, Zea, Avena, Hordeum, Secale, Triticum, Sorghum, Picea, Caco, and Populus.

As used herein, “transgenic plant” includes reference to a plant that comprises within its genome a heterologous polynucleotide. Generally, the heterologous polynucleotide is stably integrated within the genome such that the polynucleotide is passed on to successive generations. The heterologous polynucleotide may be integrated into the genome alone or as part of a recombinant expression cassette. “Transgenic” is used herein to include any cell, cell line, callus, tissue, plant part or plant, the genotype of which has been altered by the presence of heterologous nucleic acid including those transgenics initially so altered as well as those created by sexual crosses or asexual propagation from the initial transgenic. The term “transgenic” as used herein does not encompass the alteration of the genome (chromosomal or extra-chromosomal) by conventional plant breeding methods or by naturally occurring events such as random cross-fertilization, nonrecombinant viral infection, nonrecombinant bacterial transformation, nonrecombinant transposition, or spontaneous mutation.

The sequences of the present invention can be used to transform or transfect any plant. In this manner, genetically modified plants, plant cells, plant tissue, seed, and the like can be obtained. Transformation protocols as well as protocols for introducing nucleotide sequences into plants may vary depending on the type of plant or plant cell, i.e., monocot or dicot, targeted for transformation. Suitable methods of introducing nucleotide sequences into plant cells and subsequent insertion into the plant genome include microinjection (Crossway et al. (1986) Biotechniques 4:320-334), electroporation (Riggs et al. (1986) Proc. Natl. Acad. Sci. USA 83:5602-5606, Agrobacterium-mediated transformation (Townsend et al., U.S. Pat. No. 5,563,055), direct gene transfer (Paszkowski et al. (1984) EMBO J. 3:2717-2722), and ballistic particle acceleration (see, for example, Sanford et al., U.S. Pat. No. 4,945,050; Tomes et al. (1995) “Direct DNA Transfer into Intact Plant Cells via Microprojectile Bombardment,” in Plant Cell, Tissue, and Organ Culture: Fundamental Methods, ed. Gamborg and Phillips (Springer-Verlag, Berlin); and McCabe et al. (1988) Biotechnology 6:923-926). Also see Weissinger et al. (1988) Ann. Rev. Genet. 22:421-477; Sanford et al. (1987) Particulate Science and Technology 5:27-37 (onion); Christou et al. (1988) Plant Physiol. 87:671-674 (soybean); McCabe et al. (1988) Bio/Technology 6:923-926 (soybean); Finer and McMullen (1991) In Vitro Cell Dev. Biol. 27P:175-182 (soybean); Singh et al. (1998) Theor. Appl. Genet. 96:319-324 (soybean); Datta et al. (1990) Biotechnology 8:736-740 (rice); Klein et al. (1988) Proc. Natl. Acad. Sci. USA 85:4305-4309 (maize); Klein et al. (1988) Biotechnology 6:559-563 (maize); Tomes, U.S. Pat. No. 5,240,855; Buising et al., U.S. Pat. Nos. 5,322,783 and 5,324,646; Tomes et al. (1995) “Direct DNA Transfer into Intact Plant Cells via Microprojectile Bombardment,” in Plant Cell, Tissue, and Organ Culture: Fundamental Methods, ed. Gamborg (Springer-Verlag, Berlin) (maize); Klein et al. (1988) Plant Physiol. 91:440-444 (maize); Fromm et al. (1990) Biotechnology 8:833-839 (maize); Hooykaas-Van Slogteren et al. (1984) Nature ( London ) 311:763-764; Bowen et al., U.S. Pat. No. 5,736,369 (cereals); Bytebier et al. (1987) Proc. Natl. Acad. Sci. USA 84:5345-5349 (Liliaceae); De Wet et al. (1985) in The Experimental Manipulation of Ovule Tissues, ed. Chapman et al. (Longman, New York), pp. 197-209 (pollen); Kaeppler et al. (1990) Plant Cell Reports 9:415-418 and Kaeppler et al. (1992) Theor. Appl. Genet. 84:560-566 (whisker-mediated transformation); D'Halluin et al. (1992) Plant Cell 4:1495-1505 (electroporation); Li et al. (1993) Plant Cell Reports 12:250-255 and Christou and Ford (1995) Annals of Botany 75 :407-413 (rice); Osjoda et al. (1996) Nature Biotechnology 14:745-750 (maize via Agrobacterium tumefaciens ); all of which are herein incorporated by reference.

The modified plant may be grown into plants in accordance with conventional ways. See, for example, McCormick et al. (1986) Plant Cell. Reports 5:81-84. These plants may then be grown, and either pollinated with the same transformed strain or different strains, and the resulting hybrid having the desired phenotypic characteristic identified. Two or more generations may be grown to ensure that the subject phenotypic characteristic is stably maintained and inherited and then seeds harvested to ensure the desired phenotype or other property has been achieved.

The degradative enzymes can be fermented in a bacterial host and the resulting bacteria processed and used as a microbial spray. Any suitable microorganism can be used for this purpose. See, for example, Gaertner et al. (1993) in Advanced Engineered Pesticides, Kim (Ed.).

The genes of the invention can be introduced into microorganisms that multiply in plants (epiphytes) to deliver enzymes to potential target crops. Epiphytes can be gram-positive or gram-negative bacteria, or fungi, for example.

The microorganisms that have been genetically altered to contain at least one degradative gene and protein may be used for protecting agricultural crops and products. In one aspect of the invention, whole, i.e., unlysed, cells of the transformed organism are treated with reagents that prolong the activity of the enzyme produced in the cell when the cell is applied to the environment of a target plant. A secretion signal sequence may be used in combination with the gene of interest such that the resulting enzyme is secreted outside the host cell for presentation to the target plant.

It may be preferable to provide for secretion of certain of the enzymes while providing for others to remain in the cytoplasm or targeted to organelles such as, for example, the chloroplast. Generally, the first two gene products, the carboxylesterase and the flavin amine oxidase will be secreted. Secretion leaders are known in the art. Plant signal sequences, including, but not limited to, signal-peptide encoding DNA/RNA sequences which target proteins to the extracellular matrix of the plant cell (Dratewka-Kos et al., (1989) J. Biol. Chem. 264:4896-4900), the Nicotiana plumbaginifolia extension gene (DeLoose, et al. (1991) Gene 99:95-100), signal peptides which target proteins to the vacuole like the sweet potato sporamin gene (Matsuka et al. (1991) PNAS 88:834) and the barley lectin gene (Wilkins et al. (1990) Plant Cell 2:301-313), signal peptides which cause proteins to be secreted such as that of PRIb (Lind et al. (1992) Plant Mol. Biol. 18:47-53), or the barley alpha amylase (BAA) (Rahmatullah et al. (1989) Plant Mol. Biol. 12:119) and hereby incorporated by reference, or from the present invention the signal peptide from the ESP1 or BEST1 gene, or signal peptides which target proteins to the plastids such as that of rapeseed enoyl-Acp reductase (Verwaert et al. (1994) Plant Mol. Biol. 26:189-202) are useful in the invention. Such secretion signal sequences can be used in expression cassettes to provide for secretion of the protein of interest outside the cell.

The remaining gene products are provided in the cell. These enzymes include a flavin monooxygenase, an aldehyde dehydrogenase and an alcohol dehydrogenase. The expression cassettes for the corresponding genes may contain an organellar targeting sequence. In plants, for example, a chloroplast targeting sequence may be used.

In this manner, at least one of the genes encoding a fumonisin-degradation enzyme of the invention may be introduced via a suitable vector into a microbial host, and said transformed host applied to the environment or plants or animals. Microorganism hosts that are known to occupy the “phytosphere” (phylloplane, phyllosphere, rhizosphere, and/or rhizoplane) of one or more crops of interest may be selected for transformation. These microorganisms are selected so as to be capable of successfully competing in the particular environment with the wild-type microorganisms, to provide for stable maintenance and expression of the gene expressing the polypeptide, and, desirably, to provide for improved protection of the enzymes of the invention from environmental degradation and inactivation.

Such microorganisms include bacteria, algae, and fungi. Of particular interest are microorganisms, such as bacteria, e.g., Pseudomonas, Erwinia, Serratia, Klebsiella, Xanthomonas, Streptomyces, Rhizobium, Rhodopseudomonas, Methylius, Agrobacterium, Acetobacter, Lactobacillus, Arthrobacter, Azotobacter, Leuconostoc, and Alcaligenes; fungi, particularly yeast, e.g., Saccharomyces, Pichia, Cryptococcus, Kluyveromyces, Sporobolomyces, Rhodotorula, and Aureobasidium. Of particular interest are such phytosphere bacterial species as Pseudomonas syringae, Pseudomonas fluorescens, Serratia marcescens, Acetobacter xylinum, Agrobacteria, Rhodopseudomonas spheroides, Xanthomonas campestris, Rhizobium melioti, Alcaligenes entrophus, Clavibacter xyli, and Azotobacter vinlandii; and phytosphere yeast species such as Rhodotorula rubra, R. glutinis, R. marina, R. aurantiaca, Cryptococcus albidus, C. diffluens, C. laurentii, Saccharomyces rosei, S. pretoriensis, S. cerevisiae, Sporobolomyces rosues, S. odorus, Kluyveromyces veronae, and Aureobasidium pullulans.

Illustrative prokaryotes, both Gram-negative and -positive, include Enterobacteriaceae, such as Escherichia, Erwinia, Shigella, Salmonella, and Proteus; Bacillaceae; Rhizobiaceae, such as Rhizobium; Spirillaceae, such as photobacterium, Zymomonas, Serratia, Aeromonas, Vibrio, Desulfovibrio, Spiritlum; Lactobacillaceae; Pseudomonadaceae, such as Pseudomonas and Acetobacter; Azotobacteraceae; and Nitrobacteraceae. Among eukaryotes are fungi, such as Phycomycetes and Ascomycetes, which includes yeast, such as Saccharomyces and Schizosaccharomyces; and Basidiomycetes yeast, such as Rhodotorula, Aureobasidium, Sporobolomyces, Pichia and the like.

Characteristics of particular interest in selecting a host cell for purposes of production include ease of introducing the protein gene into the host, availability of expression systems, efficiency of expression, stability of the protein in the host, and the presence of auxiliary genetic capabilities. Other considerations include ease of formulation and handling, economics, storage stability, fermentation suitability, and the like.

A number of ways are available for introducing a gene expressing the degradation enzyme into the microorganism host under conditions that allow for stable maintenance and expression of the gene. For example, expression cassettes can be constructed that include the DNA constructs of interest operably linked with the transcriptional and translational regulatory signals for expression of the DNA constructs, and a DNA sequence homologous with a sequence in the host organism, whereby integration will occur, and/or a replication system that is functional in the host, whereby integration or stable maintenance will occur.

Transcriptional and translational regulatory signals include but are not limited to promoter, transcriptional initiation start site, operators, activators, enhancers, other regulatory elements, ribosomal binding sites, an initiation codon, termination signals, and the like. See, for example, U.S. Pat. Nos. 5,039,523 and 4,853,331; EPO 0480762A2; Sambrook et al. supra; Maniatis et al., eds. (1982) Molecular Cloning: A Laboratory Manual (Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.); Davis et al., eds. (1980) Advanced Bacterial Genetics (Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.); and the references cited therein.

It is recognized that the construction of a catabolic pathway in a transformed organism is a complicated feat. Therefore, any means for assembling the enzymes of interest into an organism of interest is encompassed. For example, a single polynucleotide sequence encoding all of the desired enzymes or multiples thereof may be transformed into the host organism. When microorganisms are to be applied to the environment or to a plant, several microorganisms, each transformed with one, two, three, or more nucleotide sequences of the invention, may be utilized. In this manner, all of the enzymes necessary to bring about detoxification of fumonisin and related products may be presented to the environment or to the plant by applying a mixture of transformed organisms or a single organism capable of expressing the entire pathway or at least expressing enough of the pathway to detoxify fumonisin.

In plants, nucleotide sequences for an enzyme may be transformed into a plant and crossed with plants expressing a different enzyme. In this manner, progeny can be obtained having the entire sequence or enough of the sequence to detoxify fumonisin. Alternatively, a plant can be transformed with nucleotides encoding several enzymes at the same time. In some tissue culture systems it is possible to transform callus with one nucleotide sequence, establish a stable culture line, and then transform the callus a second time with a second nucleotide sequence. The process may be repeated to introduce additional sequences.

To facilitate the expression of more than one enzyme in a cell, e.g. a plant cell, fusion proteins may be created. Generally, a spacer region is included between the proteins. The spacer region may comprise a cleavage site for cleavage by an endogenous or introduced protease.

The present invention also relates to a method of detoxifying a fumonisin or a structurally related mycotoxin with the enzymes from Bacterium 2412.1 during the processing of grain for animal or human food consumption, during the processing of plant material for silage, or food crops contaminated with a toxin-producing microbe, such as but not limited to, tomato. Since the atmospheric ammoniation of corn has proven to be an ineffective method of detoxification (see Haumann (1995) INFORM 6:248-257), such a methodology during processing is particularly critical where transgenic detoxification is not applicable.

In this embodiment, the fumonisin degradative enzymes found in Bacterium 2412.1, are presented to grain, plant material for silage, or a contaminated food crop, or during the processing procedure, at the appropriate stages of the procedure and in amounts effective for detoxification of fumonisins and structurally related mycotoxins. Detoxification by this method can occur not only during the processing, but also any time prior to or during the feeding of the grain or plant material to an animal or incorporation of the grain or food crop into a human food product, or before or during ingestion of the food crop.

The active ingredients of the present invention are normally applied in the form of compositions and can be applied to the crop area or plant to be treated, simultaneously or in succession, with other compounds. These compounds can be both fertilizers or micronutrient donors or other preparations that influence plant growth. They can also be selective herbicides, insecticides, fungicides, bactericides, nematicides, mollusicides, or mixtures of several of these preparations, if desired, together with further agriculturally acceptable carriers, surfactants, or application-promoting adjuvants customarily employed in the art of formulation. Suitable carriers and adjuvants can be solid or liquid and correspond to the substances ordinarily employed in formulation technology, e.g., natural or regenerated mineral substances, solvents, dispersants, wetting agents, tackifiers, binders, or fertilizers.

The enzymes can be introduced during processing in appropriate manners, for example as a wash or spray, or in dried or lyophilized form or powered form, depending upon the nature of the milling process and/or the stage of processing at which the enzymatic treatment is carried out. See generally, Hoseney (1990) Principles of Cereal Science and Technology (American Association of Cereal Chemists, Inc.) especially Chapters 5, 6, and 7; Jones (1992) Food Safety (Eagan Press, St. Paul, Minn.) especially Chapters 7 and 9; and Jelen (1985) Introduction to Food Processing (Restan Publishing Company, Reston, Va.). Processed grain or silage to be used for animal feed can be treated with an effective amount of the enzymes in the form of an inoculant or probiotic additive, for example, or in any form recognized by those skilled in the art for use in animal feed. The enzymes of the present invention are expected to be particularly useful in detoxification during processing and/or in animal feed prior to its use, since the enzymes display relatively broad ranges of pH activity. The esterase enzymes from Exophiala spinifera, American Type Culture Collection Accession No. 74269, showed a range of activity from about pH 3 to about pH 7 (U.S. Pat. No. 5,716,820, supra). The APAO enzyme from Exophiala spinifera, American Type Culture Collection Accession No. 74269 has a pH range of activity from pH 5 to pH 10. While not limited thereto, it is expected that the enzymes of the present invention will exhibit similar activity.

In another embodiment, ruminal microorganisms can be genetically engineered to contain and express at least one of the fumonisin degradation enzymes of the invention. The genetic engineering of microorganisms is now an art-recognized technique, and ruminant microorganisms so engineered can be added to feed in any art recognized manner, for example as a probiotic or inoculant. In addition, microorganisms, plants, or other organisms or their cultured cells in vitro capable of functioning as bioreactors can be engineered so as to be capable of mass producing the fumonisin degrading enzymes of the invention. Cite use of Bacterium 2412.1 itself, engineered to express these enzymes constitutively by manipulating regulatory factors, e.g. fccJ (LysR homolog).

Another embodiment of the present invention is the use of the enzymes of the present invention as detection reagents for fumonisins and related compounds. The enzymes of the present invention can be used as detection reagents because of the high specificity of the esterase and deaminase enzymes, and the fact that hydrolysis followed by amine oxidation can be monitored by detection of hydrogen peroxide or ammonia using standard reagents (analogous to a glucose detection assay using glucose oxidase). Hydrogen peroxide is often measured by linking a hydrogen peroxide-dependent peroxidase reaction to a colored or otherwise detectable peroxidase product (e.g., Demmano et al. (1996) European Journal of Biochemistry 238(3):785-789). Ammonia can be measured using ion-specific electrodes: Fritsche et al. (1991) Analytica Chimica Acta 244(2):179-182; West et al. (1992) Analytical Chemistry 64(5):533-540, and all herein incorporated by reference) or by GC or other chromatographic method.

For example, recombinant or non-recombinant, active fumonisin esterase, APAO, and proteins of the invention are added in catalytic amounts to a sample tube containing an unknown amount of fumonisins (FB1, FB2, FB3, FB4, or partial or complete hydrolysis products of these). The tube is incubated under pH and temperature conditions sufficient to convert any fumonisin in the sample to AP1, the AP1 to 2-OP, ammonia, and hydrogen peroxide, and to further degradation products. Then suitable reagents are added for quantification of the hydrogen peroxide or ammonia that were generated stoichiometrically from fumonisins. By comparison with control tubes that received no esterase or APAO enzyme, the amount of fumonisin present can be calculated in direct molar proportion to the hydrogen peroxide or ammonia detected, relative to a standard curve.

This invention can be better understood by reference to the following nonlimiting examples. It will be appreciated by those skilled in the art that other embodiments of the invention may be practiced without departing from the spirit and the scope of the invention as herein disclosed and claimed.

EXPERIMENTAL

EXAMPLE 1

Bacterial isolates

Bacterium 2412.1 isolates from maize were isolated as described in U.S. Pat. No. 5,716,820 and pending U.S. application Ser. Nos. 08/888,949 and 08/888,950, both filed Jul. 7, 1997, and herein incorporated by reference.

Alkaline hydrolysis of FB1 to AP1

FB1 or crude fumonisin C 8 material was suspended in water at 10-100 mg/ml and added to an equal volume of 4 N NaOH in a screw-cap tube. The tube was sealed and incubated at 60° C. for 1 hr. The hydrolysate was cooled to room temperature and mixed with an equal volume of ethyl acetate, centrifuged at 1000 RCF for 5 minute and the organic (upper) layer recovered. The pooled ethyl acetate layers from two successive extractions were dried under N 2 and resuspended in distilled H 2 O. The resulting material (the aminopentol of FB1 or “AP1” was analyzed by TLC.

Analysis of Fumonisins and Metabolism Products

Analytical thin-layer chromatography was carried out on 100% silanized C 18 silica plates (Sigma #T-7020; 10×10 cm; 0.1 mm thick) by a modification of the published method of Rottinghaus (Rottinghaus et al. (1992) J. Vet. Diagn. Invest. 4:326, and herein incorporated by reference).

To analyze fumonisin esterase activity, sample lanes were pre-wet with methanol to facilitate sample application. After application of from 0.1 to 2 μl of aqueous sample, the plates were air-dried and developed in MeOH:4% KCl (3:2) or MeOH;0.2 M KOH (3:2) and then sprayed successively with 0.1 M sodium borate (pH 9.5) and fluorescamine (0.4 mg/ml in acetonitrile). Plates were air-dried and viewed under long-wave UV.

For analysis of APAO activity, an alternative method was used. Equal volumes of sample and 14 C-AP1 (1 mg/ml, 50 mM Na-phosphate pH 8) substrate were incubated at room temperature for six days. Analytical thin-layer chromatography was then carried out on C60 HPK silica gel plates (Whatman #4807-700; 10×10 cm; 0.2 mm thick). After application of from 0.1 to 2 μl of aqueous sample, the plates were air dried and developed in CHCl 3 :MeOH:CH 3 COOH:H 2 O (55:36:8:1). Plates were then air dried, and exposed to PhosphorImager screen or autoradiographic film. A Storm PhosphorImager was used to scan the image produced on the screen.

EXAMPLE 2

Isolation of Fumonisin-Catabolizing Gene Cluster

A FB1 catabolizing esterase was identified in the culture supernatant of Bacterium 2412.1, and a gene (BacEst) was cloned from this bacterium. The gene exhibited fumonisin esterase activity similar to that of the black yeast esterase ESP1. Since FB1 is catabolized to CO 2 in this bacterium, other proteins involved in the breakdown and transport of fumonisin would also be present in induced cells of Bacterium 2412.1.

The identification and isolation of a fumonisin detoxification cluster in Bacterium 2412.1 has been accomplished. The BacEst and CTP genes as well as other structural genes and at least 3 regulatory genes have been cloned. Sequence analysis indicates that these genes code for a set of proteins that are involved in the further catabolism and transmembrane transport of fumonisin and its catabolic products (see Table 1 and FIG. 1 ). These proteins provide a catabolic pathway for the degradation of fumonisins, and could also be used to engineer a fumonisin-regulated gene transcription system.

In order to identify fumonisin catabolic genes other than bacterial esterase, it was noted that many bacterial pathway genes are clustered together on the bacterial chromosome often containing one or more operons. Both structural and regulatory genes that regulate transcription of individual pathway genes often are found in such clusters. A 4.3 kb genomic fragment from a lambda Zap II phage library, which contained a complete, functional fumonisin esterase gene (BacEst) and in addition a complete open reading frame coding for a citrate transport protein (CTP), predicted to be involved in transport of tricarballylate into the bacterium was cloned. Therefore, approximately 500 bp DNA subclones from the distal ends of this 4.3 kb fragment was used as 32 P-radiolabeled probes to identify additional lambda clones from this portion of the bacterial chromosome (i.e. chromosome “walking”). The probes were designated as P-BacEst (SEQ ID NO: 54) (at the 5′ end relative to BacEst direction of transcription), and P-CTP (SEQ ID NO: 55) (at the 3′ end). A genomic library used earlier to clone BacEst (Sau3A partially digested DNA from 2412.1, size selected for 4 to 10 kb insert size, cloned into lambda Zap II from Strategene) was used. A series of overlapping clones was identified by plaque hybridization using both P-BacEST and P-CTP probes together, and these clones were characterized by PCR using combinations of primers from probes and from the vector flanking regions. An overlapping series of clones representing various amounts of sequence extending out from the original 4.3 kb fragment was chosen based on a novel PCR screening method in which phage lysates were subjected to PCR with vector primers and probe-specific primers, in order to gauge the degree of overlap and extension. Phagemids chosen in this way were excised from lambda phage, plasmid DNA prepared from standard plasmid mini-preps and sequenced. For a second round of walking, distal probes (SEQ ID NOS: 56 and 57) were prepared from both ends of the longest 5′ and 3′ sequence obtained in the first round and library screening was repeated.

A 100 mL culture of bacterium 2412.1 was grown to saturation in LB broth and high molecular weight genomic DNA was isolated using standard methods (Short Protocols in Molecular Biology, 2 nd Ed., pp.2-11). The genomic DNA was digested with BamHI and dephosphorylated with Shrimp Alkaline Phosphatase.

The SuperCos I Cosmid Vector Kit (Stratagene) was used for library construction. The SCosI vector was digested with XbaI, dephosphorylated with Shrimp Alkaline Phosphatase, then digested with BamHI. The genomic DNA was ligated into the SCosI vector then packaged into XLI-Blue MR E. coli cells. The primary cosmid library was titered at 50 cfu/uL with a total of 30,000 cfu's. Cosmid DNA was prepared from eight colonies and digested with NotI to excise the inserts to estimate insert size. Average insert size was 27 kb with a range of 17.5-35kb. The primary library was amplified and titered at 2×10 8 cfu/mL.

Probes were made from the ends of the current BAM gene cluster and designated SCos — 5′ (SEQ ID NO: 58) and SCos — 3′ (SEQ ID NO: 59). The library was screened with 50,000 cfu's/plate using the SCos — 5′ probe and yielded 35 clones which hybridized to the SCos — 5′ probe. Inserts from these clones were excised using NotI which also produced smaller fragments of DNA from each clone. The fragments were then subcloned into the pSPORTI vector (GibcoBRL) that was previously digested with NotI and dephosphorylated.

The pSPORTI fragments were transformed into E. coli DH5a cells, DNA was miniprepped and digested with NotI to verify subclones with the correct inserts. The subclones were then sequenced with M13F and M13R primers to obtain DNA sequence from the ends of each subclone. Primer-walking was then used to fully sequence each subclone. The 8.5 kb subclone contigged with the 5′end of the BAM gene cluster.

The individual lambda clones and cosmids were sequenced to the point of overlap, and a single large contig assembled from the various clones (FIG. 2 ). Table II summarizes the American Type Culture Collection deposits made containing the overlapping clones of the fumonisin catabolic gene cluster. The strain designation and their corresponding plasmids are provided.

TABLE 2
STRAIN DESIGNATION	FRAGMENT	PLASMID
SuperCos_8.5_1	SCos 8.5	PSPORT1:
		SuperCos_8.5_1
BAM_PL1	BAM 4.3	PBK-CMV:BAM 4.3
	BAM 7.4	PBK-CMV:BAM 7.4
	BAM 8.4	PBK-CMV:BAM 8.4
	BAM 10.2	PBK-CMV:BAM 10.2
	BAM 11.1	PBK-CMV:BAM 11.1
	BAM 17.3	PBK-CMV:BAM 17.3
	BAM 3.1	PBK-CMV:BAM 3.1.1
BAM_PL2	REG 6.3	PBK-CMV:REG 6.3.1
	REG 14.3	PBK-CMV:REG 14.3.1
	REG 14.1	PBK-CMV:REG 14.1.1
	REG 8.2	PBK-CMV:REG 8.2.1
	REG 3.1	PBK-CMV:REG 3.1.1
	FMO 12.3	PBK-CMV:FMO 12.3.1
	FMO 12.2	PBK-CMV:FMO 12.2.1
	FMO 7.1	PBK-CMV:FMO 7.1.1
	FMO 5.1	PBK-CMV:FMO 5.1.1
	FMO 11.1	PBK-CMV:FMO 11.1.1

The contig represents 24.5 kb of chromosomal DNA, and contains twenty open reading frames including the fumonisin esterase and citrate transport homolog. Most of the genes in this region have homology to genes known to function in carbon catabolic pathways that involve functional groups that would be expected to be utilized for fumonisin. These have been given appropriate designations as fccA through fccT (fumonisin catabolic cluster) (Table 1). One open reading frame, fccJ, has homology to negative regulatory genes controlling lysine degradation pathway (LysR) and could be the regulator that prevents transcription of genes in this cluster in the absence of substrate. Other open reading frames in this region may be involved in other cellular functions needed for effective catabolism of fumonisin. A map of the open reading frames in the fumonisin catabolic gene cluster of Bacterium 2412.1 is shown in FIG. 3 .

Many of the gene functions deduced from ORFs in this cluster have also been identified among AP1-induced transcripts from Exophiala spinifera (see U.S. application Ser. Nos. 08/888,949; 08/888,950; and Table 2). These are, aside from the fumonisin esterase: flavin monooxygenase, aldehyde dehydrogenase, alcohol dehydrogenase, and transmembrane permease. A possible degradation pathway for fumonisins based on these enzyme activities is presented in FIG. 1 . The functional similarity of bacterial fcc cluster genes and AP1-induced Exophiala cDNAs increases the likelihood that these genes are involved in fumonisin breakdown in the respective organisms.

To identify an open reading frame corresponding to a deaminating enzyme such as the AP1-specific amine oxidase identified as an induced transcript in E. spinifera (APAO) cosmid clones corresponding to flanking regions of the fcc cluster have been obtained. It is anticipated such a gene will be found in this flanking DNA. However, identification of this gene is not crucial to this invention, as we have demonstrated AP1 deaminating activity of the Exophiala spinifera cDNA for APAO in a bacterial expression system, so it is possible to engineer the fungal gene along with bacterial genes in this cluster to engineer a complete pathway.

cDNA sequences for a citrate/tricarballylate transport protein (fccB) (SEQ ID NO: 4); a flavin monooxygenase (fccC) (SEQ ID NO: 6); an aldehyde dehydrogenase (fccD) (SEQ ID NO: 8); an alcohol dehydrogenase (fccE) (SEQ ID NO: 10); a CoA ligase (fccF) (SEQ ID NO: 12); an acetohydroxyacid synthase (fccG) (SEQ ID NO: 14); a vitamin B12 receptor (fccH) (SEQ ID NO: 16); a permease (fccI) (SEQ ID NO: 18); and a regulatory protein (fccJ) (SEQ ID NO: 20); a fumarate reductase/aspartate oxidase (fccK) (SEQ ID NO: 22); a TonB dependent receptor (fccL) (SEQ ID NO: 24); a protein with homology to N-methyl transferase (fccM) (SEQ ID NO: 26); a citrate utilization B (fccN) (SEQ ID NO: 28); a carbohydrate regulatory gene (fccO) (SEQ ID NO: 30); a possible flavoenzyme (fccP) (SEQ ID NO: 32); a leucine regulatory protein homolog (fccQ) (SEQ ID NO: 34); fccR (SEQ ID NO:36); fccS (SEQ ID NO: 38); fccT (SEQ ID NO: 40) have been isolated. Such sequences are provided. An esterase (fccA) also known as BacEst has previously been described.

The complete 24.5 kb contig of SEQ ID NO: 1 is likely to code for all the regulatory, transport, and catabolic machinery needed to catabolize FB1 to nontoxic metabolites. A single full-length cosmid clone that contains all of these genes together in order to test this hypothesis. This can readily be done by screening a cosmid library with probes generated from sequence on both ends of the contig, and selecting clones that hybridized under stringent conditions to both probes. The cosmid-containing E. coli strain would then be evaluated for its ability to break down fumonisin or AP1 added to the culture medium.

EXAMPLE 3

Pichia Expression of Degradative Enzymes

For cloning into Pichia pastoris expression vector, pPicZalphaA, oligonucleotide primers can be designed that contain a 22 bp overlap of the 5′ end (sense strand) and 3′ end (antisense strand), respectively of the open reading frame from Bacterium 2412.1, including the stop codon. In addition, each oligo has a 5′ extension with digestible restriction sites that allows cloning of the amplified insert in-frame both into EcoRI/Notl digested pPicZalphaA. pPicZalphaA is an E. coli compatible Pichia expression vector containing a functional yeast alpha factor secretion signal and peptide processing sites, allowing high efficiency, inducible secretion into the culture medium of Pichia.

Pichia can be transformed as described in Invitrogen Manual, Easy Select™ Pichia Expression Kit, Version B, #161219, with the enzyme polynucleotide of interest with either an intron added (negative control, no expression) or an intron not added (capable of making an active protein). The Pichia culture fluids and pellets are assayed for enzyme activity as described earlier.

The sample 50 UL cell pellets are resuspended in 150 UL cold 50 mM Na-phosphate, pH8.0 and divided into two fresh 500 UL tubes. One tube is kept on ice with no treatment, the pellet suspension, and one tube is used for lysis. An equal volume of 0.1 mm zirconia-silica beads is added to each tube. The tubes are BeadBeat™ for 15 seconds then cooled on ice 5 minutes. This is repeated three times. The crude lysate is then transferred to another tube for assay or lysate suspension.

The TLC assays are performed as follows:

samples:

1.) pellet suspensions (“PELL”); 10 uL

2.) lysate suspensions (“LYS”); 10 uL

3.) media controls-mixed 5 uL media with 5 uL crude bacterial enzyme (if available); 10 uL

4.) positive control-used crude bacterial enzyme (if available) undiluted; 10 uL

5.) substrate control-used 50 mM Na-phosphate, pH8.0; 10 uL

a cofactor (if required) is added to each reaction mixture

incubate 10 uL each sample+10 uL 14 C-substrate (fumonisin, metabolite, or other potential substrate) (1 mg/mL, pH8) at room temperature

spot 1.0 uL onto C18 and C60 TLC plates

develop C18 plates in MeOH:4% KCl (3:2)

develop C60 plates in CHCl 3 :MeOH:CH 3 COOH:H 2 O (55:36:8:1)

air-dry plates

expose plates to PhosphorScreen 2-3 days

use Storm Phosphorlmager (Molecular Dynamics) to develop images

EXAMPLE 4

Expression of Enzymes in E. coli

The vector for expressing the enzyme of interest is a prokaryotic glutathione S-transferase (GST) fusion vector for inducible, high-level intracellular expression of genes or gene fragments as fusions with Schistosoma japonicum GST. GST gene fusion vectors include the following features: a lac promoter for inducible, high-level expression; an internal lac I q gene for use in any E. coli host; and the thrombin factor Xa or PreScission Protease recognition sites for cleaving the desired protein from the fusion product. The insert of interest may be cloned into the 5′ EcoRI site and a 3′ NotI site allowing in-frame expression of the fusion peptide. Generation of such an insert is described in the previous example.

E. coli was transformed with the vector containing the coding sequence for the enzyme as described in BRL catalogue, Life Technologies, Inc., catalogue; Hanahan (1983) J. Mol. Biol. 166:557; Jessee et al. (1984) J. Focus 6:4; King et al. (1986) Focus 8:1, and hereby incorporated by reference. The transformed E. coli may be induced by addition of IPTG (isopropyl b-D-thiogalactopyranoside) for expression of the polypeptide of interest.

EXAMPLE 5

Transformation and Regeneration of Transgenic Plants

Immature maize embryos from greenhouse donor plants are bombarded with a plasmid containing the fumonisin-degradation/transporter enzyme nucleotide sequences operably linked to a ubiquitin promoter (FIG. 4 ). This plasmid also contains the selectable marker gene PAT (Wohlleben et al. (1988) Gene 70:25-37) that confers resistance to the herbicide Bialaphos. The preferred construct for expression in maize is the nucleotide sequence of the degradative enzyme either fused to the barley alpha amylase signal sequence or organellar targeting sequence, or left intact for expression in the cytoplasm. Transformation is performed as follows. All media recipes are in the Appendix.

Preparation of Target Tissue

The ears are surface sterilized in 30% Chlorox bleach plus 0.5% Micro detergent for 20 minutes, and rinsed two times with sterile water. The immature embryos are excised and placed embryo axis side down (scutellum side up), 25 embryos per plate, on 560Y medium for 4 hours and then aligned within the 2.5-cm target zone in preparation for bombardment.

Preparation of DNA

A plasmid vector comprising the fumonisin-degradation/transporter enzyme operably linked to the ubiquitin promoter is made. This plasmid DNA also contains a PAT selectable marker. The plasmid is precipitated onto 1.1 μm (average diameter) tungsten pellets using a CaCl 2 precipitation procedure as follows:

100 μl prepared tungsten particles in water

10 μl (1 μg) DNA in TrisEDTA buffer (1 μg total)

100 μl 2.5 M CaCl 2

10 μl 0.1 M spermidine

Each reagent is added sequentially to the tungsten particle suspension, while maintained on the multitube vortexer. The final mixture is sonicated briefly and allowed to incubate under constant vortexing for 10 minutes. After the precipitation period, the tubes are centrifuged briefly, liquid removed, washed with 500 ml 100% ethanol, and centrifuged for 30 seconds. Again the liquid is removed, and 105 μl 100% ethanol is added to the final tungsten particle pellet. For particle gun bombardment, the tungsten/DNA particles are briefly sonicated and 10 μl spotted onto the center of each macrocarrier and allowed to dry about 2 minutes before bombardment.

Particle Gun Treatment

The sample plates are bombarded at level #4 in particle gun #HE34-1 or #HE34-2. All samples receive a single shot at 650 PSI, with a total of ten aliquots taken from each tube of prepared particles/DNA.

Subsequent Treatment

Following bombardment, the embryos are kept on 560Y medium for 2 days, then transferred to 560R selection medium containing 3 mg/liter Bialaphos, and subcultured every 2 weeks. After approximately 10 weeks of selection, selection-resistant callus clones are transferred to 288J medium to initiate plant regeneration. Following somatic embryo maturation (2-4 weeks), well-developed somatic embryos are transferred to medium for germination and transferred to the lighted culture room. Approximately 7-10 days later, developing plantlets are transferred to 272V hormone-free medium in tubes for 7-10 days until plantlets are well established. Plants are then transferred to inserts in flats (equivalent to 2.5″ pot) containing potting soil and grown for 1 week in a growth chamber, subsequently grown an additional 1-2 weeks in the greenhouse, then transferred to classic 600 pots (1.6 gallon) and grown to maturity. Plants are monitored for the expression of a fumonisin-degrading/transporter protein.

APPENDIX

APPENDIX
272 V
	Ingredient	Amount	Unit
	D-I H 2 O	950.000	Ml
	MS Salts (GIBCO 11117-074)	4.300	G
	Myo-Inositol	0.100	G
	MS Vitamins Stock Solution ##	5.000	Ml
	Sucrose	40.000	G
	Bacto-Agar @	6.000	G

Directions:

@=Add after bringing up to volume

Dissolve ingredients in polished D-I H 2 O in sequence

Adjust to pH 5.6

Bring up to volume with polished D-I H 2 O after adjusting pH

Sterilize and cool to 60° C.

##=Dissolve 0.100 g of Nicotinic Acid; 0.020 g of Thiamine.HCL; 0.100 g of Pyridoxine.HCL; and 0.400 g of Glycine in 875.00 ml of polished D-I H 2 O in sequence. Bring up to volume with polished D-I H 2 O. Make in 400 ml portions. Thiamine.HCL & Pyridoxine.HCL are in Dark Desiccator. Store for one month, unless contamination or precipitation occurs, then make fresh stock.

Total Volume (L)=1.00

288 J
	Ingredient	Amount	Unit
	D-I H 2 O	950.000	Ml
	MS Salts	4.300	g
	Myo-Inositol	0.100	g
	MS Vitamins Stock Solution ##	5.000	ml
	Zeatin .5 mg/ml	1.000	ml
	Sucrose	60.000	g
	Gelrite @	3.000	g
	Indoleacetic Acid 0.5 mg/ml #	2.000	ml
	0.1 mM Abscisic Acid	1.000	ml
	Bialaphos 1 mg/ml #	3.000	ml

Directions:

@=Add after bringing up to volume

Dissolve ingredients in polished D-I H 2 O in sequence

Adjust to pH 5.6

Bring up to volume with polished D-I H 2 O after adjusting pH

Sterilize and cool to 60° C.

Add 3.5g/L of Gelrite for cell biology.

##=Dissolve 0.100 g of Nicotinic Acid; 0.020 g of Thiamine.HCL; 0.100 g of Pyridoxine.HCL; and 0.400 g of Glycine in 875.00 ml of polished D-I H 2 O in sequence. Bring up to volume with polished D-I H 2 O. Make in 400 ml portions. Thiamine.HCL & Pyridoxine.HCL are in Dark Desiccator. Store for one month, unless contamination or precipitation occurs, then make fresh stock.

Total Volume (L)=1.00

560 R
Ingredient	Amount	Unit
D-I Water, Filtered	950.000	ml
CHU (N6) Basal Salts (SIGMA C-1416)	4.000	g
Eriksson's Vitamin Mix (1000X SIGMA-1511)	1.000	ml
Thiamine.HCL 0.4 mg/ml	1.250	ml
Sucrose	30.000	g
2,4-D 0.5 mg/ml	4.000	ml
Gelrite @	3.000	g
Silver Nitrate 2 mg/ml #	0.425	ml
Bialaphos 1 mg/ml #	3.000	ml

Directions:

@=Add after bringing up to volume

#=Add after sterilizing and cooling to temp.

Dissolve ingredients in D-I H 2 O in sequence

Adjust to pH 5.8 with KOH

Bring up to volume with D-I H 2 O

Sterilize and cool to room temp.

Total Volume (L)=1.00

560 Y
Ingredient	Amount	Unit
D-I Water, Filtered	950.000	ml
CHU (N6) Basal Salts (SIGMA C-1416)	4.000	g
Eriksson's Vitamin Mix (1000X SIGMA-1511)	1.000	ml
Thiamine.HCL 0.4 mg/ml	1.250	ml
Sucrose	120.000	g
2,4-D 0.5 mg/ml	2.000	ml
L-Proline	2.880	g
Gelrite @	2.000	g
Silver Nitrate 2 mg/ml #	4.250	ml

Directions:

@=Add after bringing up to volume

#=Add after sterilizing and cooling to temp.

Dissolve ingredients in D-I H 2 O in sequence

Adjust to pH 5.8 with KOH

Bring up to volume with D-I H 2 O

Sterilize and cool to room temp.

**Autoclave less time because of increased sucrose**

Total Volume (L)=1.00

All publications and patent applications mentioned in the specification are indicative of the level of those skilled in the art to which this invention pertains. All publications and patent applications are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be obvious that certain changes and modifications may be practiced within the scope of the appended claims.

59 1 24494 DNA Bacterium 2412.1 Fumonisin Catabolic Gene Cluster 1ggccgcaatt aaccctcact aaagggatct tattggccat acggcgatag ttagcggcga 60gcgcgcccag gtcgacgcgc aggtggccgc ccgccatgtc cgaagtcgtg tcgttcacca 120tcgtccagcc cccgcttttc gcgctcgctg aagcctaggt gtgacttgtt cgaatatcag 180gcctatctat cgcttagtct gcggatttat ccgcactact aaaatataat gctgaggtta 240cgccacatga tgccagcgct ggacgcgatc gaccgcaaga tcattggcct gcttcgggtc 300aatggccgca tgcccaacaa tgagttggcg cagaaggtag ggctttcgcc ttccgcctgt 360ctgcgacgcg tcaagttgct cgagtcgaac ggggtgatcc gagggtattg tgcattggtt 420gccgagcagt cgttggacgc cagcgtggtg gcgatcgtcc ggataacctt ggacaagcag 480accgaggact atctgaatcg gttcgaggag gccgttcggc ggcatcccga gatcgctgag 540tgctttctga tgaccggcga cgcagactac atccttcggg ctaccgcgcc gagcacggcc 600gcctacgagc aaatccacaa ggaagtcctt tctcggcttc ccggggtggc gcgcatccat 660tcgagcttcg ccatccgcag cgtgctgtcg tcggtcgcaa ggccctagag ctagccgcgt 720tcggccacgg cggctcgacg agctcgagac ctaatgcacg attaatatcg aatatcccac 780tctgccgctc gaccgacgcg gcggcgtcgc agctcgcgcg cagcgcaggc ccggctgggt 840gcacattatt caagcacggg tgcgtcacct gaatggcgtc gcggcacgat tcaagaggtg 900aactcgataa atattgaata aggcgccggt ctgccggagc atgggagtgc aaagacactg 960gcagcagggg atctgaccat gcctagggat acgacaccga atttcgctcc ggtcaccacg 1020gcaaaagagg gccgccgaca ccgaggcagc accgccttac gaaggctcat gctgacggcg 1080gccggcagcg ccctggtgct gggtcttgcg cccaaggcgc tcgcgcaggt ggcggttccg 1140ccggctggtc acgaggcgtc gcaggaggtg caggagatcg tcgtcaccgc gcagcgccgc 1200agcgagaaca ttcagaatgt gccggtctcg gtgcaggcgc tgtcggcagc gcagctcgag 1260cgcgaaggga tcaaacagac cagcgatatc gcccgagtga cgcccaacgt caccatcgcc 1320atgcccaacg gcgaaggcaa ccagccggcg gtgacgatcc gcggcatcgg cctcaacgac 1380ttcaattcca acaacgccgg cccgaacgcg atctatgtcg acgatgtcta tatcagcgcc 1440ccgtcggccc agaccttcgg aatcttcgac atcaaccaga tccaggttct caaaggaccg 1500caaggtacgc tctatgggcg caactccagc ggtggggcct tggtgttcac gtccagagcg 1560ccgagccaag acttcgccgc ggacgcccat ttcgattacg gcagctacaa cacctatcaa 1620ctgcaagccg gcgtcggcgg ccctctgagc gatcagctaa gcgcccgcct ggccttcgtc 1680gtcaaccact ccgacgggtt catgcacaac acgctgacgg gcggttcggc gtcgggcacg 1740gacaatcagg ccgtcaggct gcaactgctc taccgaccta atgacaggct gaaagtactt 1800ctcagttcgg cctatggtca tgtcaactcg ccgatcgtcc agtaccgaca cttgggcgcc 1860ttcgcggcag gaacccaatc cagcgccagc ccgactctct gcagccccga gcaggtccgc 1920gccggaggtt gcgtcaacgt gttcggcgca ggcacgccga gcggcttcta cgacggttcc 1980agcgatcgcg gtgaacgctt gcgcgtggaa aacttcctgc agcaggcccg cgccgactat 2040gaggtcggtc cggtgaccct gacatcgatc agcgccttca cgcacagcaa aaagagcggc 2100cccgacgacg ccgacgggac gtctgacagt ctgctccacg cgacctacgg cgttcgctcc 2160gacacctgga cccaagagtt ccgcgccgcc tattccggcc agcgcctgca ttgggtggcg 2220ggcgcctact atctcgacga gaccctcaag caaaatcagc cacttagcat cttctacgat 2280ggagatcgct tcggcggcct gggcatcccg gccagggcgg gagccttcga cggcatcgcg 2340caaaagagct taagccaaaa cactcagaaa acacggtcga tagccgcctt cggccaagcc 2400gactatacct tggaccggtt caccctgacc ttgggcggtc gttacaccca tgaacgcaag 2460acgttcgatc acttcagcgc gacccaggtc caagcaggag gccttgggaa atacggtcct 2520ctcggcaaga tcgtctcgct gagcgaagcg ttcaaggctt ccgatccgac ctggcgcgcc 2580gcgctttcct accgtcccgc cgagcgtgtt atggtctacg gcagcgtcgc caccggcttt 2640aagggcggcg ccttcaacgg cgggttcctg agcagcaacc ccaacaaagc cctcgccgcg 2700gtcaaacccg tcgcaccgga gaaggtgacc acctacgaac tgggcttcaa gtcgagcctg 2760ttcgagcgtc gcctggtggt caacggcgcg gctttctaca acagctacga caacgagcag 2820atcctggcca acacggccgt cgtcgtggat accgtgaccg gccctgttac cgtgacgacg 2880aacgtcctga ccaacgcccg aaaggcccac tcccagggcg tggaattgga agtaaaggcc 2940gtcccgatcc cggatctcgt cctcagcctg cagccggcct ggctgcgaac gcggctggac 3000gaggcgggct tctccggggg aacgtcgctg gaaggcaagc aactggccaa tgcgccgaag 3060ttctcgctct acgccgcggc ggactacacc ttccatcttg ccgacgacga cagcgtcaac 3120gtcgccttca cctcggccta caagtcgcac cagttcttcg attcgacgaa cgccccctat 3180acccagcagg agggctactg ggtgcacaac gccagcctga ccttcaactc cagaaaccac 3240tgggatgtcg ggttcaatgt ccgaaacctg acgggcacga agtactacaa ctatctgttc 3300gacgaggggg cgacgttcgg cttcatcaac ggcgtcgtgg ccgcgccgcg gacctacagc 3360gtgcaattca acctgcatct ctaggcgcgc agaaggaggc gccgccatgc gcggcgcctc 3420ccgctatcgg actgaaacgt taggtcctct tccccgcttc ggcgcgcggc gtcgccgcgc 3480gccgtctctc tttgcaaccg tccgcggcga cccttgctgt cgcctagcag cgcggcgcgc 3540tccatccatg cgccatgcgc aatgaatgcg acgatcgttg catttgatgg aggtttcgga 3600ggccgctagt tctgctgcga gggggctccc gaccatgggc gccgtagccg cggctggcgc 3660ggttcgcccc ctcgactgtc ccgacgtcgt cggcgcggac cgtagcgatc cgatcggaag 3720gaatgggtgc gaagttgaag tatgacgtgg tggtggtcgg gggcggcaat gcggcgatga 3780cggcggccgt caccgcgcgg gaagccggcg cgacggtgct ggtgcttgag catgcgcccc 3840ggtcgatgcg cggcggcaac agccgccata cgcgcaacat gcgcacgatg cacgaggcgc 3900cacttgcggt cttgaccggg caatattccg aagacgaata ctggaacgac ctgaagcggg 3960tcacgggcgg ggaaaccgac gaggccctgg cccgtctggt gatccgcagc acgacggacg 4020ccatcccctt catgctccgg tgcggcgtgc gcttccagcc atcgctgtcg ggcaccttga 4080gcctgtcgcg gaccaacgcg ttcttcctgg ggggcggcaa ggctctggtg aacgcctact 4140acgcgaccgc cgagcgcctg ggcgtcgaca tcctctatga cagcgaagtc accgagatcg 4200tgctcgaagg cggccgggtc cggcgtctgg tggtccgcag ccaggggttc cccatcgagg 4260tggaggcgcg cgcggtgatc gcctcgtcgg gcggcttcca ggccaacctg caatggctgg 4320cgaacgcctg gggcccggcg gcgtcgaatt tcatcgtacg cgggacgccc tacgcgacgg 4380gcacggtgct gcgcaacctg ctcgaccagg gcgtggcctc ggtgggcgat ccgacccagt 4440gccatgctgt cgccatcgac gggcgcgcgc ccaagtacga cggggggatc gtcacccgac 4500tggactgcgt gccgttctcg atcgtggtca atcgcgacgg ccaacgcttc tacgacgagg 4560gcgaggacat ctggcccaag cgatatgcga tctgggggcg tctgaccgcg caacagcccg 4620atcagatcgc ctacagcatc atcgacagcc gatccgaacg acttttcatg ccgtcggtgt 4680ttcccccgat caaagccgac tcgatttccg aactcgcggc caagctcggg ctggagccgg 4740cgacgctcgc gcagaccatc gagacgttca atcgcgcctg ccaacccggt cgcttcgatc 4800cgcaggatct tgacggggtc cgcaccgagg ggatcacgcc gtgcaagtcc aattgggccc 4860ggccgatcac cgagccgccg ttcagcgcat atcccctgcg gcccggcatc accttcacct 4920acctcggcgt caaggtcgat gaacgcgcca gggtgatcct ggcctccggc cagccgacag 4980agaacctgtt cgcgtctggc gagatcatgg ccgggagcat tcttgggcgc ggttacctgg 5040cgggcttcgg catggcgatc gggaccgtct tcggacgcat tgcgggccgg gaggccgcat 5100atcatgcagc ataatgtcct ggatttcgtg accaagacgc gcacgggcga gccgcgcccg 5160gccgaaacgc ccgcgatcat cgaagcgcgc cggaccatgg aggtttgcaa cgcctgtcgc 5220tattgcgaag gctactgcgc ggtctttccg gccatgaccc tcaagcggga gttcgaggaa 5280gccgatctca cctacctggc caatctctgt cactcgtgcc gcggctgtta ctacgcttgc 5340caatacgcgc cgccccatga gttcgggatc aacgtgccca aggtgctggc cgaggtccgc 5400accgaaagct accaggccca tgcctggccg caggccgtcg ccgtcgcctt cgagcgtaac 5460ggtctggtgg tgtccctgag cgctgcactc gcgatcgttg tcgtgctgct gggaacggcc 5520ttcttcaatg gatcggcgat gttccaggcg cacgcctcga cgcccggcgc aggcttctac 5580aaggccgtgc cctatgcggt catggtgagc gtcgccggcg cgatcttcgc ctatgccgcc 5640ttggcgatgt tcatcggcct tatccggttt tggaagaccg tgggccttgg cttgcgcgac 5700gccgtcgaac cgcgaacctt gttccaggcg ctgaaggatg cggcgaccct gcgctatctc 5760ggcgggggcg gcgatggctg caacgacgtc gacgctagct tctcgacctc acgccgacgt 5820ttccatcacg ccatggccta cggcttcctg ctctgttttg cctccacctc caccggtacg 5880gtctacgacc acctcctggg ctggcccgcg ccctatccct tcttcagcct gccggtgctg 5940ctgggaacgg tcggcggaat tgggatcgtc atcggcacgc tcggactgct ctggctgaag 6000ctggtcggcg accaggagcc taggtcgaag gcgcaattgg gcgccgacac cgcgctgctg 6060gtgctgctgt tcctgatcag cgtgacgggg ctgttgctgc tggcgcttcg gacgacggcg 6120gccatgggcg tgatcctgac cgtgcacctt ggactggtct tctcgttctt cgcgacgatg 6180ccgtacagca agttcgtgca cggactctat cgaaccgtcg ccttggttcg ttacgccgtc 6240gagcgcaagg cgctggcctc cgggacgacg gaggaagcgt cttgatcttg gccnacgctt 6300tggacgccgc aggccgcgcc cgccgctcag ccgaggggcg cctgggcctt gatcgcgcat 6360cgaggggcct tggcccttgt tccgaccaat gtgtcggcgc ccgcaccgcc gtggccgaag 6420acaggagaca atgaatgaaa gcggccattt accggcgcgg ggagatcgtt gtcgataccg 6480ttcccgatcc ggttccagga ccaggccagg ttctcgtccg gagccttgtt tgcggggtat 6540gtggttcgga tctgcattac cgacatcacg cacaccggtt cgtcgatctg gccttgcgct 6600cgggcgcgcc cgccctggcc gccgatttgg atcgcgatat cgtccttggt cacgaattca 6660gcgctcaagt cgtcgactac gggcctaaga ccgagcgtct cctgaagtcg ggaacggtcg 6720tctgctcgcc ccccgtcgcg ttcggggcca gcggcatgcg cgccgttggc tactccgacg 6780aattaccggg cgggtttggc cagtacatgg tcttgaatga ggcgttcctg atgccggccc 6840caaacggact ggatccggct cgcgcggcgc tcaccgagcc gatggcggtg gggtggcacg 6900cggtgaagct ggccggtccc ggacgcgacc atatcccgct cgtgatcggc tgcgggcccg 6960tgggcatggc ggtcatcgcc gcgctccggg gtctgggcgt cggaccgatc atcgcggccg 7020acttcaatcc ggcgcgtcgg agcctggcgg cgcgcatggg cgccgatatt gtcatcgacc 7080cggcggagcg gtccccctac gacgaatggc gggataccgc ggcggcgtca ggcctggccg 7140gactggcggg ggcgccagcg tcgctgcgga cctgtctggt cttcgagtgt gtcggcctgc 7200caggaatgct gcgtcagatc atggaaggcg ccccggcgga gtcggagatc atcgtcgtcg 7260gggcctgcat ggagcccgat agcctcgagc cgatgatggc gatgcataag gctctgacgc 7320tgaattttcg cgaacctaca cgatcgagga gttcgccgag gtccttcgga tgatcggtga 7380gggcgagctc cacgtcgagc cgttgctcag ccaacccatc ggcctggaag accttccggg 7440ggtcttcgac naagcgcccg ggagggccgg gggcgccaag gtcctcgtcg acccctggcg 7500ctgacgccgc cgaaaaccaa cgacagaaaa caacgggcgg gaggaacaat tgggtagcga 7560tgagcaagac gatcctctcg tcggcggcgg tcgtcagttg cttctcaagc aatgtctgct 7620caacgtcgtt gaccaggcgg ggagcaacga gatcgtctat caaggccggc tgaggttcag 7680ctacgccgac ctgctttctc ggatttcgcg actggccgat gctttgacgg gcttgggcgt 7740caagccgggc gacaccgtgg cggttctgga gtgggacagt caccggtacc tggaatgctt 7800cttcgccatt cccatgcttg gcgccgtcat ccagacggtg aacatccgac tagcccgaga 7860cgacctgcgc tacacgctcg agcatgcggg cgccaccctg gcgctgagcc acaccgattt 7920cctgccgatc ctcgaggagg tgatcgacca attgcccagc ctgcgcgggg tcgtccatct 7980gaaggacgac gaggcggaag ccgcccatcc ctgggtgctg ggggagtatg aggccctgat 8040ggcggccgcg cgccctcggt tcgacttccc ggacttcgac gagaacacgc gggcgacgac 8100cttctacacc agcggcacga ccgggcgtcc gaagggcgtc tactattcgc atcgtcagct 8160ggtgctgcac accctggcgg tgatggcgac gctggccctt ggagacggtt acgccaggct 8220ggggcgcgat acggtctaca tgccgatcac cccgatgttc catgctcatg cgtggggaat 8280gcccttcgtg gcgacgatgg tcggctgcaa gcaagtctac ccagggcgct atgttcccga 8340gcaactggtg gagcttcagc gcgcggagaa ggtgaccttc tctcattgcg tgcccacact 8400tttgcagatg atgctcaatt cgccttcggg ccagacggcg gatttcaccg gatggcaggt 8460gctcgtcggc ggagcggcgc tgccccgcgg cctggctctt caggccgcgg ggcgcggcat 8520cgtcctgacc accggatacg gaatgtccga aaccgggccg ctggtcagct tcacgcgcat 8580taggaccgaa gcaatggctc cagctcagga ggaggtcgcc attcgcacca aggtcggaca 8640agctatcgcg ctggtcgacc tccgggtcgt ggatgagtcc atggcggatg tgccccgcga 8700cggcctctcc gcgggcgaga tcgtgttgcg tgcgccttgg ctgacggctg ggtaccatcg 8760cgatctggcc gcctcgcgcg agctttggcg cggaggaagc cttcatacgc aggatttcgg 8820ccggattgac gcggagggct acctgcagat cagcgaccgc ctccagggag tcatcaagac 8880ggtggggatg ggttctcctg agctgggaga tctcgtcagc cgccatccgg cggtgctgga 8940gagcgccgcg atcgctgtcg ccgacgagcg ttggggagag cgcccagcga tggtcgtcgt 9000gctcaggccg ggcatgagcg cgaccacggc ggacatccga gaccaccttt catcgtatgt 9060cgcgaccggc gaaatacctc gctacgccgt gcccgagcag atctggttcg tcgaggagct 9120cgaccgaacg agcgtgggca aggtcgacaa gcgggcgctt cgttccaggt tcgccgaagc 9180ggcgtcctga gatggcgcat gaccgaacgg gaccaaatgg cgtcggcgtt ggttggctct 9240ttccaaccct gttacggtcg ggtctgatga tggcgacgcg gtggtcgaac cgcgggaatg 9300ggatcgtgta gccgggatgt tgaactgcga tctattggat ctgcgcgcct tcgtcgcggt 9360ccacgaaacc cgcagcttca tccgcgcggc gcatctgctc ggcctttccc agcccgcgct 9420cagtcgccgg atccaacgct tggagggact ggtcggcggc gctctcttcg accgaaccag 9480ccggaccatg accgagaccg cgcttggcaa ggagctgctg ccggtggccc gccgaacgct 9540tgagtttctg gacaattcgc tgttcgcctc gcccaagctg cgcgaaccgc gctggaccga 9600catcagcatt ttttgcgtgc agaccgccgc gttccgcgtt ctgccgcgcg cggcccggcg 9660cttcatggat gaaaatcccc gactgcgcct gaggatcatc gatgttccgg ctgtcgaagg 9720cgcggaactg gtggcgcgag gggaagcgga gttcggtatc agcatcgaga gcctgcttcc 9780gtccggcctg cgtttcgagg ctcttcacga ggacccgttt ggcttggcgt gccatcggag 9840ccatcgcctg gcgcaaagcg acgtcatcga atggtccgcg ctccgcggcg aaaatcttgt 9900cgccgtccac cgggccagtc gcaaccggac cctgctcgac gccgagctca agcagcatgc 9960gatctccctg gactggcgtt acgaggtcgg tcacttgacg accgcgctgg ggctgatcga 10020gtccgaggtc ggcgtggccg tcatgccgcg gatggtgatg ccccaatcag gccgctcaga 10080actggtctgg gttcccttgg tcgccccggt cgtgaggcgc acgatcggca tcgtgcagcg 10140ccgggtgggc gcgatgcatc ccgccgccgc ccaactgctc gagcggttgc gggaggaatg 10200gccgaccggc gcgcccgcgg acgagtagac cgccaagatt cgaggtccgt gcgggcgcgc 10260ggcggctggc cggtcgtcac cgcggccccg tcaccgcgtc atattcagca cctgcacttg 10320ggtgaactcg tgcagccctt cctcgcctaa ctccgagccc atccccgaga acttggcgcc 10380gccgagcggg agatcgggct gcacgtcggc gtgcttgttc acccagaccg agccggcctc 10440catatcggcg gccaggctcc acgcgcggac gacgttgcgg gaccagatgg atccccccag 10500accatagggc gaggcgttgg cgcggcgcac ggcgtcgacg gggtcggagt accggatcac 10560cggcatcacc gggccaaact gctcttcgtc gacgagctga gcgccttcgg cgatgtcgcg 10620tacgatggtg ggctcgatga aatagccctt gtcgcccttg cggcggccgc cggcgatgat 10680gcggccgtcc gtcctcgcac gctcgattag accaagaacc ttctcgaact ggcgccggtt 10740ctgcagcggc cccatctgaa cgccctgttc gagtccatcg cccacgaccg tgcgggccgc 10800caactgcgcg aactcctcgc acatggcctc atagaggctc tcatggacgt aaatccgttt 10860ggcggcgatg cacacctgac cggcgttttg catggccgcg gcgaacaccc tgggagcgac 10920ttccttgggg tcgacgtcat ccaggacgat cagagcgtcg tttccgccca actcaagcga 10980tatacgtttg aggccttcgg ccgcgccggc catgaccttt tttccggtct gggtcgatcc 11040ggtgaagctg attttgcgaa tgccaggatg gcgggtcatt tccgcgccga gatcgtcggc 11100gtcggtgatg atgttaatga cgcccggtgg gacgatatcc ttgaccaagg cgccaaaccg 11160aagcgccgtc agaggcgtcg tcgccgccgg cttgaggatg accgtgttgc cggccagcag 11220ggccgccggg atcttgaacg ccatcaacag catcgggaaa ttccagggga cgatgcagcc 11280caccacgcct agggggcgtc tatgcacctc tacgcggccc gtcgcgtcgt ctctgaccac 11340gcgaggcggc agatcgagcg aggtgaagta gcggaagaag gccgcggagg cgtagatctc 11400gcccatcgcg tctgcgagcg gcttgccctg ttcctgcgtc agcaaacgcg ccagcgccga 11460ctggtcggct tcaattgcgt cggcgatggc gttgagcgtg gccctgcgct gctcgagcgt 11520ggttgcccgc cagctctgaa aggcgcgttc cgcggcggcg acggcttcgt ccagttggtc 11580gcggtcggcc ctggggcaat cgatcaccag cggcgtttca gtcgcgggat tgattacgga 11640catcgtcgtt gcgccggcga ccaggcggcc gtcgatcagc agcttgtact ctagcatgtt 11700ggctccgatg gtcggctgat cgagccgccg gtttgcatgc ggcgctcgat cacgggattg 11760gagatcgact tcaaccggtc aggccctgat agccctcgtg gaagcgcatg gcgatcgacg 11820ggggaaaggc caggttcgca gccctatctg tccggacccg cacaagggcc ggtccgagcg 11880catcgcgcgc cgcgctcaga gcgggccgca gatccggcgc ggtctcggcc ttgaagccga 11940agcatcccag gctttcggcc aacctttccc agaggacggg gcccatctcg gtgccgaagg 12000tcctgccgta gcgcgcctgc tcattcggga cctccatcga ccacgaccct tcggccagga 12060cgacgacggt caccttgacg tcctcgcgga cggcggactg cagttccata caatggaagc 12120cggccgcgcc gtcgccagta acgcagacga tctggcgatc cgggcttccg agacccgcgc 12180cgatcgcaga cggtatgccg gtgcccagca tccccatctc aagaatgttc aggtacgagc 12240gcggtctggt cgagggcaac ataaagtgag cccaaaggct cgtgaaaccg ccatcggcga 12300cgtacaccgc atcgggaccg aaaacctctc ccaccgtctg cataacctca gccgggtgag 12360gacttggacc gccatgggcc tcgatatgag cgaactcgga acgccgccat tcggcgtcca 12420tttggcgata gcgcgtgagg tcgacgtccg cgcccgagcg cgtgggcgcg ttctcgaggg 12480cctcgaggag gccttccacg aggctgcccg cgtccgaaac gatgcctagc gtcaacggac 12540gtgaggcgcc caaattgcgg gggtcgatat cgacctggat cagcttgtgc ccctccgatg 12600agccccaata gcggtcgaag ggcgtgtcga tgtttcccag gcgcgtgccc agcgccagga 12660tgacgtcggc ttcgcgtcgc gcctgatcag cgccggcgcc ataggcgtgc aggtggaggg 12720ggtgatcttg agggacggcc gaccggcccg ccaggctggc gatgacgccg caacccagct 12780tgtcggctag gcgcagcaca gcctcgcccg cgccagctcg gtcgaccccg gagccgacca 12840tgatcagcgg acgagtcgcg gcggccagca attgggcggc ggcgttgatt tgcgagccgc 12900cggctgaagg agggggcgcc cgataggcga tcggatcgag caggccagcc cgggactcgt 12960cggtcatgtc gtacatcact gggctcggaa catcgatctg gaccgggccg ggacgccccg 13020cccacatctc ccggaacgcc atgcgcgtaa cctcgccgat gcgttgccag gtatgaatgg 13080gcgcgcccca tttgaccgcc gggcgcagga gttccaattg gtcggcgccc tggaacgtgc 13140tgggcgtcgc gggatagacg accccgccat gatgctgcgc ggtgatggcg atcatcggca 13200cgccctcatg cttggcggtc accaagccgg gcagcaggtt tgccgtaccg ggtcctgggt 13260ttgtcacggt ggcggcgact tggcctgtcg tcttgtagag accctcggcc atataggccg 13320cagccgcctc atgccgcacg gggatgaagc ggatgccgtt gtcgtcgagc gccgccagca 13380gcggatcaac ctccggcgac atcaagccga aaacaaaccg cacgccttcg cgcgctaggc 13440actgcgcaag taatgcgccc ccggtgatcg ccatttgttt ctccctgcgt acgacggcgc 13500atggaacacc gccgcgtctg gtcgttgtcc gaccacgttg ccgacttttc gaactatcag 13560tcgataatta tcgaaacagc aaccggtctc gtttgcaata cgagccgtag gccgccattc 13620tgatcgatga gaaacgaagt ggcgcgatgc ggttcaaacg cttgttttat ggggcgtttc 13680gccatcgcga catgcgttcg gcgcattgat cgccccgatc attgcattgg ctggcggact 13740ggcgcgccga tagtcgttgc gatggtcgcg agaataagcg tgcgaagtgg gaggatgtga 13800agatgggggc caggagtatg tgtgcgggac ggttcggacg cttctgcatt ggcttggctt 13860catcggttgc cgtgactcta gggggagcct ccgccgccgg cgcggcaacc gcgacggatt 13920ttccggtccg caggaccgat ctgggccagg ttcagggact ggccggggac gtgatgagct 13980ttcgcggaat accctatgca gcgccgccgg tgggcgggct gcgttggaag ccgccccaac 14040acgcccggcc ctgggcgggc gttcgccccg ccacccaatt tggctccgac tgcttcggcg 14100cggcctatct tcgcaaaggc agcctcgccc ccggcgtgag cgaggactgt ctttacctca 14160acgtatgggc gccgtcaggc gctaaacccg gccagtaccc cgtcatggtc tgggtctacg 14220gcggcggctt cgccggcggc acggccgcca tgccctacta cgacggcgag gcgcttgcgc 14280gacagggcgt cgtcgtggtg acgtttaact atcggacgaa catcctgggc tttttcgccc 14340atcctggtct ctcgcgcgag agccccaccg gaacttcggg caactacggc ctactcgaca 14400ttctcgccgc tcttcggtgg gtgcagagca acgcccgcgc cttcggaggg gaccccggcc 14460gagtgacggt ctttggtgaa tcggccggag cgagcgcgat cggacttctg ctcacctcgc 14520cgctgagcaa gggtctcttc cgtggcgcta tcctcgaaag tccagggctg acgcgaccgc 14580tcgcgacgct cgccgacagc gccgcctcgg gcgagcgcct cgacgccgat ctttcgcgac 14640tgcgctcgac cgacccagcc accctgatgg cgcgcgccga cgcggcccgc ccggcatcgc 14700gggacctgcg caggccgcgt ccgaccggac cgatcgtcga tggccatgtg ctgccgcaga 14760ccgacagcgc ggcgatcgcg gcggggcagc tggcgccggt tcgggtcctg atcggaacca 14820atgccgacga aggccgcgcc ttcctcgggc gcgcgccgat ggagacgcca gcggactacc 14880aagcctatct ggaggcgcag tttggcgacc aagccgccgc cgtggcggcg tgctatcccc 14940tcgacggccg ggccacgccc aaggaaatgg tcgcgcgcat cttcggcgac aatcagttca 15000atcggggggt ctcggccttc tcggaagcgc ttgtgcgcca gggcgcgccc gtgtggcgtt 15060atcagttcaa cggtaatacc gagggtggaa gagcgccggc tacccacgga gccgaaattc 15120cctacgtttt cggggtgttc aagctcgacg agttgggtct gttcgattgg ccgcccgagg 15180ggcccacgcc cgccgaccgt gcgctgggcc aactgatgtc ctccgcctgg gtccggttcg 15240ccaagaatgg cgaccccgcc ggggacgccc ttacctggcc tgcctattct acgggcaagt 15300cgaccatgac attcggtccc gagggccgcg cggcggtggt gtcgcccgga ccttccatcc 15360ccccttgcgc ggatggcgcc aaggcggggt gacgccgtcg acgatggcgt gacgacggtc 15420gaggcgatgt tctcgatctg gagtccgcgc cgcctcgatt tgcgtcgtct ccggcgctca 15480gacgaacgcc ccagttccat ccacacagtc actttcccga gccgagctgt cggcggtggg 15540acgaaagaga agccgatgcg agccataccg attcaaagcc gacccgttcc atccgatcgc 15600caagccccgg ctcgcggggt gacgcaatga cggccgcgga ggagcgccgc gggcatttgg 15660gtaagatcct gcgcgtggcg agcggcaact tcctcgagca gtacgacttc ttcatctacg 15720gctactacgc gacctacatc gcccaggtgt tctttccatc gggcgacgag acgacgtcgt 15780tgatgctctc cctggccacc tttggcgtcg ggttcctgat gcggccgcta ggggcgatca 15840ttctcggatc ctacatagat cgcgtcggcc gccggcaggg cttgatcgtc acgttgggga 15900tcatggcgat cggcacgctc accatcgccc tgacgccggg ttacagcgcc atcgggatcg 15960ccgcgccgct catcgtcgtc gccggtaggc tcttgcaggg cttctccgcc ggagccgaac 16020tcggcggcgt ctcgatctac ctggcggaaa tcgcaaagcc tggtcgacgg ggcttctaca 16080cctcctggca gtcggccagc cagcaggtcg cggtgatggc ggccgcgctt gtcggtctaa 16140gcctcggcgc aacattgacg cccgaccaaa tgcaccagtg gggctggcgc gttccgttgc 16200tgctcggctg tgcgatcgtc cccgtcatcc tgtggctccg ccgatcgctc gacgagaccg 16260aggcctataa acacattcat cacaaggccc attccctatt gggctcgctg gcccagttgg 16320ggggcagctg gaggccgatc ttggccggca tggcgatctc ggtcctgacc accacgacct 16380tctatatgat caccgcctac acgccgacct tcggaaagca ggctctcggt ttggacgccc 16440aggacgtcct cgtcgtcacc atgctggtcg gcgcctcgaa ctttatatgg ctgccggtcg 16500gcggcgcgct ctccgactgg attgggcgca cgccggtgct cctctctgtg cccctggtgg 16560tccttgtcgc ggcctacccg ctgatcgcct ggctggtggg cgcgccatcg ttcttcgcgt 16620tcgcgacggc gctgctggcc ttgtcggtct gctttggcct ctataacggc gcgatgatcg 16680cacgactgac cgaactgatg ccgcctgcgg cacggacgct ggggttttcc ctggcgttca 16740gtctggccac gtcgctgttc ggaggcttca cgccgctggt cagcacttat ctgatcagcg 16800cgaccggtaa caaggccgca cccgcgctgt ggctgtgctt cgccgcgatg atcagcctga 16860tcggggtatt ggcttcacgc aggatggggg ccgaccccga tcgatccatg gcttgaaggc 16920gctcgccgcc gacggcctcc cgtacccaga tcctatcggc ggagccagcc ggcctgcgga 16980tccccgacac gggcgttccg caggccggtt ttcttgttcg tggggaggag gttggcctgc 17040ggcgctcgat ctcgcgagcc atgggccgcc cccaatcttt cggctgggcc gtgcggcgtt 17100gggcgcgccg gccatttcga cggcggcttg cgccgccatc gacacgtcgt tcaggccggg 17160gctttggtgt tcgcatgtcg ggactttaaa gtagccgccc gccgccaggg gccggccacg 17220acgtagagcg ccgccgccgc aacgaggagc cggccgaaac ctggaatgcg cctcccaccc 17280gattatcgtc aatgacgaac gaggccgcca acggaccggc gctgaagccc accaacgcga 17340gaggggcgag cagtacggcg gcggtcctgc tgggctccaa ggcgatcacg tccgccacca 17400gaaacggctg cagcgccagc cagaacaggc caaagccaca agcgcttgcg atgagcgcga 17460cgggcgtgcc ggcgtgaagc aggccgatga caagaccggc ctgcagcact gcgccggcgg 17520ccagaaccgt acgggcgtgc acgcgcgcac cgagccagga tgctgcaaga gcacccgcca 17580cctggaaggc caggctgccc gcgatcgcgg cgccgaccgt ggccggggcg aaatggtgtt 17640gcgcggccag gcgctccagg tagttccatg ccgccccgat gccggcgttt tgaagaaacg 17700ccgcgagcgc cacgaccatc agggccggag agacgactac gcgaccatgg tgcgatcccg 17760taggagcagg cacgtggtcg acgatcgccg gcgcaacgag gcaggaggcc atcgcaacgc 17820ccgccaagac cgcgaacccg gcatccacgc caaaccgcgg gatcacccag atcggcagca 17880aataggccgc gatcacctgg gggatcgtgg aaagaccgag cagcagcccg ctcatgcgct 17940caggccggtc gttgtgggta aggatggcgc cggccgcgcc cagcaatagc ccttccagca 18000acccggccgc gccgcgttgg accaggatcg tggccgggga tgcggcccag tagatggcga 18060ggttgatgat cgcaaggagc agggaggcag cggccacctt ggcgcgcatg tggccgaggt 18120tcatcaggaa cggacccgcc gtcgagcctg cggccagggc gaagacttcg atcatggcgg 18180cctgacctac gcccgcctcg ctgatccgcc cggcgttggc gagacccccg agcaggatgg 18240gttcgacgcc catcaccagc atcgaggccg tgccgatcag catagtggcc gatacgacgg 18300acacggcctg gtgatggcga ggtccggtat tccggggcag ggtcagggac ataggctcca 18360actgtcgtcg agaaggtgaa gggaattcgc atctgtgggg gagcggctgc gctccacaca 18420cccagcaaat atcgataata gtttgagagc aattctgaca tcgtccgcga tttacgaaaa 18480cacgcatatt ttaattgcct gccgctggat taagcagata aatatcgagt taagcctccg 18540gtgcgttttc gatcccgaag ttggtgatca tcggaccgcc gatagtcagg gcgcacgaaa 18600ccagggaggt gagcatgccg gatggtgagg ctatcagcgg cgtcggccag tggttggcgg 18660ctttcgagag cgcgctgaaa cgcgacgatc tggcggcggc ggtcgaccta tttgtcgagg 18720acgtcttttg gcgagacatc gtcgccttca cctgggatat ccgcacgctc gaggggcgcg 18780gggcgatcca agagcttcta ggagccgcct cggggctcgc aaggtccgcg tcctggtcga 18840cgacgtcggc cgatcatgat caggaaggcg tcgtcagctt tgaaaccgac ctggggcgag 18900gccatggtta ccttcgcctc cgggggggac gctgctcgac attgctgacc tgtctggaag 18960aactcacggg acatgaggag acacgcggcc ctcgccgtcc gcgcggggcg agcgtaggcc 19020cggcggaccc tgacgaaaat tggaagaatc gtctcgaggc ggaaagccgg gcgatggggc 19080gtgagaccca gccgttcgtg cttatcgtcg gcggcggtca aggcggtcta gcgcttggcg 19140cgcgcctccg tcagctccag gtcccgactc tgatcgtcga tcagcaccca cgggtggggg 19200accaatggcg atcgcggtac gcatcgctct gcctgcacga tccagtctgg tacgaccacc 19260ttccttacct gccgtttccc gatacttggc cggtttatac gcccaaggac aagatcggcg 19320attggctcga agcttatgcg caggcgatgg agctgctggt ctggtgttcg accagatgcg 19380tgtccgccgt ctatgacgcc gaagccgggc gatggaccgt caccctgcgc cgaggcgagg 19440agaccagcgt catccgcccc gcgcatctgg tcctggcgac gggcaacgcc ggcaagccgc 19500gcgttccgcg cttcaagggc caagcgcagt tcgaaggtcc gatcctgcac tcgagcgcct 19560atcggagcgg ggctgatttc aaaggacggc gcgtggccgt gatcggatcg aacaattcgg 19620cccacgacat ctgcgcagac ctcgtggccc acggcgttga cgtcaccatg atccagcgca 19680gttcgaccca tgtcgtccgt tccgaaacgg tcatgcggac catgctcgcg ccgctttatt 19740cagaggaggc cttggcggcc ggcataggca cggagctggc cgacctgctt gtggcttcca 19800tgccgttacg cctgcaggcc gaaggctatc gcgccctcca cgtcgcgatc gccgagcagg 19860acgcagcgtt ctacgccgcg ctcgaggcga tcggcttcat gcatgacttc ggcgaggacg 19920gcaccggcat gccgctgaag tatcttcgtc gcgcgtcggg gtactatatc gacgtcggcg 19980catccgaact cctggccagc ggggccataa agctgcgctc ccgcgtcgag atcgatcact 20040tcgacaccga cggcctggcc ctctcggacg gcagcaaggt cgacgccgac gccgtcatct 20100gcgcaaccgg tttcggctcc atggacgagt gggcggccga attgatttcc cccgaggtcg 20160cggccaaggt cggaagggtc tggggctatg ggtccggcac ccgaggcgat ccgggcccct 20220gggagggcga acttcggaac atgtggaagc ccacccgcca gcagggcttg tggttccagg 20280gcggaaacct ggcgcaaacc cgcttctact ccagagcgct cgctctgcag ttgaagcccg 20340acatgctgat tgccgtgagt ctacgttcgt caccgactag gcggcggcga agctcactga 20400tcttgctccg cggcgtccca gaggctcacg ctcgcctctc cctggtctat tgcttcgcga 20460atttgcggcc gcagctcgag atgttcgggc aagaacgtcc tgcaataggc gacgctcacg 20520gtcagggcct cttcgtagta gtcctcggtg atcgcaccgt tttggacaaa cgagatcgcc 20580agaaccgcat cgacgatctg gatgttcgtg tgaaacttct tctgcggatc gcgcagaaaa 20640ggcatgtgga agatgcggtc aaggcgctga taggccgcgc gagcgacggc ttccacatat 20700tcgcgatcgg cctgccgtgt ctcgagcccg ccaaatccac ccaggaatag cttcgaagcc 20760ggcgagttgg cgttgtagta atccacgcca ttgcgcaggt cccacgccgt cagcgcctgc 20820caactactga gactcttcac cggaatgggc cggcgggtga gttgctcgaa acctttcagg 20880tgcctttgcg ccagcgccag gaaggcggcc tccttggtcg ggaagaagtg atagaccgag 20940gcgggtggaa cgccggcccg ctcggcgatc tgatagaggc cgacggcggc ggggttctcg 21000tcctgtagca gcgcctcggt cgcatccagc agggccgtgt agcgagtcag cgacgtgctg 21060cgggacggcg cgcgcgggcg aaccttcttc ccggatgcgt tccgcgggtg atggggcctg 21120agtcgtctcc atgaacgaga gtagcggcgc ggcgagcctt ggatatcggc aatgatggcg 21180ccgggttgct ctatgacgag ccgaatggcg gcgctcggcg caagtgtgcg atgtaaaaaa 21240tgtgccgcgg acgtgacgtc cgcggcacac aaccgggcga tggctagagt tgctcgtatc 21300ctacagccac ctcccggtgg acgccgccag ggaggaatgg gcgtccgggg aggggagtca 21360gtagcggaag cggagcccga cgttgaaata cctgccgcgc ggatcgtagg ctgcgctcgt 21420cgggaccgag aagctggacg gattgaccgt cgccaccggt ggatcacggt cgaagaggtt 21480gtttaccgaa gcaaaaacct gctgcttatg cccgaatgaa tcgaagctgt aggtcaccgt 21540ggcgtcagtg taccagactg cgccggtgtg gttcaggttg gtgtcgacac cttcgacgtt 21600ttcagcgtca aagaccgagc gagagatgaa gcgctcttgc aggaacagcg accatgcggc 21660gcgctcgtaa cgcgcctgca gattgagcag ccatttgggg gcggtgggtt cgccgagcga 21720ttgtaggggc gccgagccca gggcggtgac ggatgcggcc gtgcggtggt tggccaaggc 21780ccgcaggttc agagatccgc ccgcgacgtt gcgcacgtag gacgcctcca aatcgacacc 21840cgccgccttc tgcaccgcaa ggttcaggtt ggggccgatg acggtcagcg tgttgtccgc 21900attccgggtg atcaaggcgc acatggactg gttgcccgcg gcgcagaggt cgatctcctg 21960ctggggcagg aggtagtcga tggcgtcttt gagatcgatg atatagcggt ccgccgaaag 22020ctgaagcccc ggggcgaagg ccggccgcaa cacgacgccg aaggtcaggg tatcggcgcg 22080ctcggggcga agatccgggt tgccggcggt gaagaagcgg gtctgcaacg tctggccctg 22140atagacagag ttcagggtag cctgacggcc ggggtcgtag agttccacca ggctggcgcc 22200gcggatgtcg cgcgagcgag tcaggcgaaa ccgcagtcca tcgacgggct cgtagtcgcc 22260gccgaccttc caggtcgtga cgccgcccga gacgctatag tcggcatacc ggaccgcgcc 22320gttgaggttg agcgctcgac caagcgcgct gtccttgagc accggaaccc cgacctcaag 22380ataagcctcc ttgatgttgt agctcccgct gaagggaaga gggttgtaga ggttgaaacc 22440accaggccgg ttgctctgag aggccggcgc tccacgcagt ccggcggtcg acgtgatcgc 22500ctgcgagatg gcgtcggtgg tctggttggc tttctcctca cgatattcgc cgcccgcggc 22560gatcgagatc ggcccggcgc ccagcgaaaa cgcctggcca aggtcgccga cgaggttcag 22620gcccgcgacg acctgttcca gtttcaggtt cgccaccccg tcgtcgagga catagtcgat 22680cgcggcggca cttggcgcgc cggcgccgaa aatgttgagc ggcacgcagc ccgcgtcgag 22740acccgaaagg gtcgaacggc agacgatctt tcccgtcggg tccttcacgg cgtcgaccgc 22800tgcatagaga ttgcggttga tcgacaggtt gttttcgcga agctccaggt tcgtgcggcc 22860gtaggagatc gaaccgtcga gcttccaggt gtcgttgagg tccgcccgga agccggcagc 22920tccgcgacgc accttggcgt aggactcgat ctcgaccagc gggaactcgc cggcgaaccg 22980gccgacggaa accgacgtca gccggttggt gtccatcagt gcgccgagtg cggtcgggag 23040gaacgcgttg tcgcggaaga tcgtgaaggc gttcgcgctg ccgacaaact ggttgacgaa 23100ggcgccgagg ttggtgtggc tataggcata ggtgccttcc gcatagagct tgacgcgttc 23160tgaggcctcg aactcgccgc ggaggaagcc attgtagcgc cgctggtccg gagcgaagcc 23220gagattgacg cgcggaccgt cgccgccgct ctggaaggaa ctgctggtaa agcttccgta 23280gttaaaggtc gcaagcgtac cgccgggaag aaaggtgacg cccttcagcg gacccgatgt 23340gatcaggccg ccgtaggcgc cgcgcgagct tcggatgtcg ggaaccaccg tgacgcccgt 23400cggggcgccg ggcacgggat attgccccgc cgcgcgatca taccacgccc gatcggtggc 23460ctgatcggcg cgaatgccgt cctcgtgata gtattcgacc gcggcaagga gatgtgcgcg 23520cccttgggca aacgacttgc cggccgccag cgatccgccc accgaggcca gatcgttgcg 23580gctcgagacg ccggtctgga cgttcgcctt taggccctcg aaatcctcgt cgaggacgaa 23640gttgattacg ccggatacgg catcggatcc gtaggcggcc gaagcgccgc cggtcaccac 23700gtcgacccgc ttgacgagag cctggggcaa cacattgacg tccaccgatc cggtgtagtt 23760ggtggcgacg aagcggttgc cgttgagcag aacgaggttt cggttcgcgc ccaggccgcg 23820caggctcagg agattctggc cgctgttgcc ggtgcccggc gtcgtgccgg ggttcgacgt 23880tttcagacta ttattgaaga ccggaagttg gttgaggcca tcggcgatgt tggttggagc 23940cgccgccttg agctgatcgc agacgccgcg gtcacgggag tcggtgcgct gaatccgctc 24000tggaggcggc tgcccgtgac gacgatttcg ctgacttcct ggcgtcctgg gccgagggtt 24060ttcgacaggt gcttgctgcg cccggccgag gcttggggct agaagcacgg acgtcgccgc 24120tgcgcccagc aggctaaggc ggaaagtgct cttctgattg gttgctgctt ggctcatggc 24180taatccctcc ctctctttgt gacggcgagg gggggacggc atggcggaac gccgcacgcg 24240aaaatgtccc ggttcctcct cgattgggcg atgtgcctcg cccttccgct atgagtaacg 24300agggcttttg ggcccatcaa atgcaatgat cggcgggctc tattcgcccg gcgcatggat 24360cttttcgatc tgccggattg gcaattcagg gttgaaaata cgataattat cgaattgagt 24420tagtgggggc tctcgtttct cgagcagaat tgatcggtgg cgtagtgagg ctgaccatgc 24480aggttgacgg gatc 24494 2 1590 DNA Bacterium 2412.1 fccA Fumonisin Esterase 2atg ggg gcc agg agt atg tgt gcg gga cgg ttc gga cgc ttc tgc att 48Met Gly Ala Arg Ser Met Cys Ala Gly Arg Phe Gly Arg Phe Cys Ile 1 5 10 15ggc ttg gct tca tcg gtt gcc gtg act cta ggg gga gcc tcc gcc gcc 96Gly Leu Ala Ser Ser Val Ala Val Thr Leu Gly Gly Ala Ser Ala Ala 20 25 30ggc gcg gca acc gcg acg gat ttt ccg gtc cgc agg acc gat ctg ggc 144Gly Ala Ala Thr Ala Thr Asp Phe Pro Val Arg Arg Thr Asp Leu Gly 35 40 45cag gtt cag gga ctg gcc ggg gac gtg atg agc ttt cgc gga ata ccc 192Gln Val Gln Gly Leu Ala Gly Asp Val Met Ser Phe Arg Gly Ile Pro 50 55 60tat gca gcg ccg ccg gtg ggc ggg ctg cgt tgg aag ccg ccc caa cac 240Tyr Ala Ala Pro Pro Val Gly Gly Leu Arg Trp Lys Pro Pro Gln His 65 70 75 80gcc cgg ccc tgg gcg ggc gtt cgc ccc gcc acc caa ttt ggc tcc gac 288Ala Arg Pro Trp Ala Gly Val Arg Pro Ala Thr Gln Phe Gly Ser Asp 85 90 95tgc ttc ggc gcg gcc tat ctt cgc aaa ggc agc ctc gcc ccc ggc gtg 336Cys Phe Gly Ala Ala Tyr Leu Arg Lys Gly Ser Leu Ala Pro Gly Val 100 105 110agc gag gac tgt ctt tac ctc aac gta tgg gcg ccg tca ggc gct aaa 384Ser Glu Asp Cys Leu Tyr Leu Asn Val Trp Ala Pro Ser Gly Ala Lys 115 120 125ccc ggc cag tac ccc gtc atg gtc tgg gtc tac ggc ggc ggc ttc gcc 432Pro Gly Gln Tyr Pro Val Met Val Trp Val Tyr Gly Gly Gly Phe Ala 130 135 140ggc ggc acg gcc gcc atg ccc tac tac gac ggc gag gcg ctt gcg cga 480Gly Gly Thr Ala Ala Met Pro Tyr Tyr Asp Gly Glu Ala Leu Ala Arg145 150 155 160cag ggc gtc gtc gtg gtg acg ttt aac tat cgg acg aac atc ctg ggc 528Gln Gly Val Val Val Val Thr Phe Asn Tyr Arg Thr Asn Ile Leu Gly 165 170 175ttt ttc gcc cat cct ggt ctc tcg cgc gag agc ccc acc gga act tcg 576Phe Phe Ala His Pro Gly Leu Ser Arg Glu Ser Pro Thr Gly Thr Ser 180 185 190ggc aac tac ggc cta ctc gac att ctc gcc gct ctt cgg tgg gtg cag 624Gly Asn Tyr Gly Leu Leu Asp Ile Leu Ala Ala Leu Arg Trp Val Gln 195 200 205agc aac gcc cgc gcc ttc gga ggg gac ccc ggc cga gtg acg gtc ttt 672Ser Asn Ala Arg Ala Phe Gly Gly Asp Pro Gly Arg Val Thr Val Phe 210 215 220ggt gaa tcg gcc gga gcg agc gcg atc gga ctt ctg ctc acc tcg ccg 720Gly Glu Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro225 230 235 240ctg agc aag ggt ctc ttc cgt ggc gct atc ctc gaa agt cca ggg ctg 768Leu Ser Lys Gly Leu Phe Arg Gly Ala Ile Leu Glu Ser Pro Gly Leu 245 250 255acg cga ccg ctc gcg acg ctc gcc gac agc gcc gcc tcg ggc gag cgc 816Thr Arg Pro Leu Ala Thr Leu Ala Asp Ser Ala Ala Ser Gly Glu Arg 260 265 270ctc gac gcc gat ctt tcg cga ctg cgc tcg acc gac cca gcc acc ctg 864Leu Asp Ala Asp Leu Ser Arg Leu Arg Ser Thr Asp Pro Ala Thr Leu 275 280 285atg gcg cgc gcc gac gcg gcc cgc ccg gca tcg cgg gac ctg cgc agg 912Met Ala Arg Ala Asp Ala Ala Arg Pro Ala Ser Arg Asp Leu Arg Arg 290 295 300ccg cgt ccg acc gga ccg atc gtc gat ggc cat gtg ctg ccg cag acc 960Pro Arg Pro Thr Gly Pro Ile Val Asp Gly His Val Leu Pro Gln Thr305 310 315 320gac agc gcg gcg atc gcg gcg ggg cag ctg gcg ccg gtt cgg gtc ctg 1008Asp Ser Ala Ala Ile Ala Ala Gly Gln Leu Ala Pro Val Arg Val Leu 325 330 335atc gga acc aat gcc gac gaa ggc cgc gcc ttc ctc ggg cgc gcg ccg 1056Ile Gly Thr Asn Ala Asp Glu Gly Arg Ala Phe Leu Gly Arg Ala Pro 340 345 350atg gag acg cca gcg gac tac caa gcc tat ctg gag gcg cag ttt ggc 1104Met Glu Thr Pro Ala Asp Tyr Gln Ala Tyr Leu Glu Ala Gln Phe Gly 355 360 365gac caa gcc gcc gcc gtg gcg gcg tgc tat ccc ctc gac ggc cgg gcc 1152Asp Gln Ala Ala Ala Val Ala Ala Cys Tyr Pro Leu Asp Gly Arg Ala 370 375 380acg ccc aag gaa atg gtc gcg cgc atc ttc ggc gac aat cag ttc aat 1200Thr Pro Lys Glu Met Val Ala Arg Ile Phe Gly Asp Asn Gln Phe Asn385 390 395 400cgg ggg gtc tcg gcc ttc tcg gaa gcg ctt gtg cgc cag ggc gcg ccc 1248Arg Gly Val Ser Ala Phe Ser Glu Ala Leu Val Arg Gln Gly Ala Pro 405 410 415gtg tgg cgt tat cag ttc aac ggt aat acc gag ggt gga aga gcg ccg 1296Val Trp Arg Tyr Gln Phe Asn Gly Asn Thr Glu Gly Gly Arg Ala Pro 420 425 430gct acc cac gga gcc gaa att ccc tac gtt ttc ggg gtg ttc aag ctc 1344Ala Thr His Gly Ala Glu Ile Pro Tyr Val Phe Gly Val Phe Lys Leu 435 440 445gac gag ttg ggt ctg ttc gat tgg ccg ccc gag ggg ccc acg ccc gcc 1392Asp Glu Leu Gly Leu Phe Asp Trp Pro Pro Glu Gly Pro Thr Pro Ala 450 455 460gac cgt gcg ctg ggc caa ctg atg tcc tcc gcc tgg gtc cgg ttc gcc 1440Asp Arg Ala Leu Gly Gln Leu Met Ser Ser Ala Trp Val Arg Phe Ala465 470 475 480aag aat ggc gac ccc gcc ggg gac gcc ctt acc tgg cct gcc tat tct 1488Lys Asn Gly Asp Pro Ala Gly Asp Ala Leu Thr Trp Pro Ala Tyr Ser 485 490 495acg ggc aag tcg acc atg aca ttc ggt ccc gag ggc cgc gcg gcg gtg 1536Thr Gly Lys Ser Thr Met Thr Phe Gly Pro Glu Gly Arg Ala Ala Val 500 505 510gtg tcg ccc gga cct tcc atc ccc cct tgc gcg gat ggc gcc aag gcg 1584Val Ser Pro Gly Pro Ser Ile Pro Pro Cys Ala Asp Gly Ala Lys Ala 515 520 525ggg tga 1590Gly 3 529 PRT Bacterium 2412.1 3Met Gly Ala Arg Ser Met Cys Ala Gly Arg Phe Gly Arg Phe Cys Ile 1 5 10 15Gly Leu Ala Ser Ser Val Ala Val Thr Leu Gly Gly Ala Ser Ala Ala 20 25 30Gly Ala Ala Thr Ala Thr Asp Phe Pro Val Arg Arg Thr Asp Leu Gly 35 40 45Gln Val Gln Gly Leu Ala Gly Asp Val Met Ser Phe Arg Gly Ile Pro 50 55 60Tyr Ala Ala Pro Pro Val Gly Gly Leu Arg Trp Lys Pro Pro Gln His 65 70 75 80Ala Arg Pro Trp Ala Gly Val Arg Pro Ala Thr Gln Phe Gly Ser Asp 85 90 95Cys Phe Gly Ala Ala Tyr Leu Arg Lys Gly Ser Leu Ala Pro Gly Val 100 105 110Ser Glu Asp Cys Leu Tyr Leu Asn Val Trp Ala Pro Ser Gly Ala Lys 115 120 125Pro Gly Gln Tyr Pro Val Met Val Trp Val Tyr Gly Gly Gly Phe Ala 130 135 140Gly Gly Thr Ala Ala Met Pro Tyr Tyr Asp Gly Glu Ala Leu Ala Arg145 150 155 160Gln Gly Val Val Val Val Thr Phe Asn Tyr Arg Thr Asn Ile Leu Gly 165 170 175Phe Phe Ala His Pro Gly Leu Ser Arg Glu Ser Pro Thr Gly Thr Ser 180 185 190Gly Asn Tyr Gly Leu Leu Asp Ile Leu Ala Ala Leu Arg Trp Val Gln 195 200 205Ser Asn Ala Arg Ala Phe Gly Gly Asp Pro Gly Arg Val Thr Val Phe 210 215 220Gly Glu Ser Ala Gly Ala Ser Ala Ile Gly Leu Leu Leu Thr Ser Pro225 230 235 240Leu Ser Lys Gly Leu Phe Arg Gly Ala Ile Leu Glu Ser Pro Gly Leu 245 250 255Thr Arg Pro Leu Ala Thr Leu Ala Asp Ser Ala Ala Ser Gly Glu Arg 260 265 270Leu Asp Ala Asp Leu Ser Arg Leu Arg Ser Thr Asp Pro Ala Thr Leu 275 280 285Met Ala Arg Ala Asp Ala Ala Arg Pro Ala Ser Arg Asp Leu Arg Arg 290 295 300Pro Arg Pro Thr Gly Pro Ile Val Asp Gly His Val Leu Pro Gln Thr305 310 315 320Asp Ser Ala Ala Ile Ala Ala Gly Gln Leu Ala Pro Val Arg Val Leu 325 330 335Ile Gly Thr Asn Ala Asp Glu Gly Arg Ala Phe Leu Gly Arg Ala Pro 340 345 350Met Glu Thr Pro Ala Asp Tyr Gln Ala Tyr Leu Glu Ala Gln Phe Gly 355 360 365Asp Gln Ala Ala Ala Val Ala Ala Cys Tyr Pro Leu Asp Gly Arg Ala 370 375 380Thr Pro Lys Glu Met Val Ala Arg Ile Phe Gly Asp Asn Gln Phe Asn385 390 395 400Arg Gly Val Ser Ala Phe Ser Glu Ala Leu Val Arg Gln Gly Ala Pro 405 410 415Val Trp Arg Tyr Gln Phe Asn Gly Asn Thr Glu Gly Gly Arg Ala Pro 420 425 430Ala Thr His Gly Ala Glu Ile Pro Tyr Val Phe Gly Val Phe Lys Leu 435 440 445Asp Glu Leu Gly Leu Phe Asp Trp Pro Pro Glu Gly Pro Thr Pro Ala 450 455 460Asp Arg Ala Leu Gly Gln Leu Met Ser Ser Ala Trp Val Arg Phe Ala465 470 475 480Lys Asn Gly Asp Pro Ala Gly Asp Ala Leu Thr Trp Pro Ala Tyr Ser 485 490 495Thr Gly Lys Ser Thr Met Thr Phe Gly Pro Glu Gly Arg Ala Ala Val 500 505 510Val Ser Pro Gly Pro Ser Ile Pro Pro Cys Ala Asp Gly Ala Lys Ala 515 520 525Gly 4 1290 DNA Bacterium 2412.1 fccB Citrate Transport nucleotide 4atg acg gcc gcg gag gag cgc cgc ggg cat ttg ggt aag atc ctg cgc 48Met Thr Ala Ala Glu Glu Arg Arg Gly His Leu Gly Lys Ile Leu Arg 1 5 10 15gtg gcg agc ggc aac ttc ctc gag cag tac gac ttc ttc atc tac ggc 96Val Ala Ser Gly Asn Phe Leu Glu Gln Tyr Asp Phe Phe Ile Tyr Gly 20 25 30tac tac gcg acc tac atc gcc cag gtg ttc ttt cca tcg ggc gac gag 144Tyr Tyr Ala Thr Tyr Ile Ala Gln Val Phe Phe Pro Ser Gly Asp Glu 35 40 45acg acg tcg ttg atg ctc tcc ctg gcc acc ttt ggc gtc ggg ttc ctg 192Thr Thr Ser Leu Met Leu Ser Leu Ala Thr Phe Gly Val Gly Phe Leu 50 55 60atg cgg ccg cta ggg gcg atc att ctc gga tcc tac ata gat cgc gtc 240Met Arg Pro Leu Gly Ala Ile Ile Leu Gly Ser Tyr Ile Asp Arg Val 65 70 75 80ggc cgc cgg cag ggc ttg atc gtc acg ttg ggg atc atg gcg atc ggc 288Gly Arg Arg Gln Gly Leu Ile Val Thr Leu Gly Ile Met Ala Ile Gly 85 90 95acg ctc acc atc gcc ctg acg ccg ggt tac agc gcc atc ggg atc gcc 336Thr Leu Thr Ile Ala Leu Thr Pro Gly Tyr Ser Ala Ile Gly Ile Ala 100 105 110gcg ccg ctc atc gtc gtc gcc ggt agg ctc ttg cag ggc ttc tcc gcc 384Ala Pro Leu Ile Val Val Ala Gly Arg Leu Leu Gln Gly Phe Ser Ala 115 120 125gga gcc gaa ctc ggc ggc gtc tcg atc tac ctg gcg gaa atc gca aag 432Gly Ala Glu Leu Gly Gly Val Ser Ile Tyr Leu Ala Glu Ile Ala Lys 130 135 140cct ggt cga cgg ggc ttc tac acc tcc tgg cag tcg gcc agc cag cag 480Pro Gly Arg Arg Gly Phe Tyr Thr Ser Trp Gln Ser Ala Ser Gln Gln145 150 155 160gtc gcg gtg atg gcg gcc gcg ctt gtc ggt cta agc ctc ggc gca aca 528Val Ala Val Met Ala Ala Ala Leu Val Gly Leu Ser Leu Gly Ala Thr 165 170 175ttg acg ccc gac caa atg cac cag tgg ggc tgg cgc gtt ccg ttg ctg 576Leu Thr Pro Asp Gln Met His Gln Trp Gly Trp Arg Val Pro Leu Leu 180 185 190ctc ggc tgt gcg atc gtc ccc gtc atc ctg tgg ctc cgc cga tcg ctc 624Leu Gly Cys Ala Ile Val Pro Val Ile Leu Trp Leu Arg Arg Ser Leu 195 200 205gac gag acc gag gcc tat aaa cac att cat cac aag gcc cat tcc cta 672Asp Glu Thr Glu Ala Tyr Lys His Ile His His Lys Ala His Ser Leu 210 215 220ttg ggc tcg ctg gcc cag ttg ggg ggc agc tgg agg ccg atc ttg gcc 720Leu Gly Ser Leu Ala Gln Leu Gly Gly Ser Trp Arg Pro Ile Leu Ala225 230 235 240ggc atg gcg atc tcg gtc ctg acc acc acg acc ttc tat atg atc acc 768Gly Met Ala Ile Ser Val Leu Thr Thr Thr Thr Phe Tyr Met Ile Thr 245 250 255gcc tac acg ccg acc ttc gga aag cag gct ctc ggt ttg gac gcc cag 816Ala Tyr Thr Pro Thr Phe Gly Lys Gln Ala Leu Gly Leu Asp Ala Gln 260 265 270gac gtc ctc gtc gtc acc atg ctg gtc ggc gcc tcg aac ttt ata tgg 864Asp Val Leu Val Val Thr Met Leu Val Gly Ala Ser Asn Phe Ile Trp 275 280 285ctg ccg gtc ggc ggc gcg ctc tcc gac tgg att ggg cgc acg ccg gtg 912Leu Pro Val Gly Gly Ala Leu Ser Asp Trp Ile Gly Arg Thr Pro Val 290 295 300ctc ctc tct gtg ccc ctg gtg gtc ctt gtc gcg gcc tac ccg ctg atc 960Leu Leu Ser Val Pro Leu Val Val Leu Val Ala Ala Tyr Pro Leu Ile305 310 315 320gcc tgg ctg gtg ggc gcg cca tcg ttc ttc gcg ttc gcg acg gcg ctg 1008Ala Trp Leu Val Gly Ala Pro Ser Phe Phe Ala Phe Ala Thr Ala Leu 325 330 335ctg gcc ttg tcg gtc tgc ttt ggc ctc tat aac ggc gcg atg atc gca 1056Leu Ala Leu Ser Val Cys Phe Gly Leu Tyr Asn Gly Ala Met Ile Ala 340 345 350cga ctg acc gaa ctg atg ccg cct gcg gca cgg acg ctg ggg ttt tcc 1104Arg Leu Thr Glu Leu Met Pro Pro Ala Ala Arg Thr Leu Gly Phe Ser 355 360 365ctg gcg ttc agt ctg gcc acg tcg ctg ttc gga ggc ttc acg ccg ctg 1152Leu Ala Phe Ser Leu Ala Thr Ser Leu Phe Gly Gly Phe Thr Pro Leu 370 375 380gtc agc act tat ctg atc agc gcg acc ggt aac aag gcc gca ccc gcg 1200Val Ser Thr Tyr Leu Ile Ser Ala Thr Gly Asn Lys Ala Ala Pro Ala385 390 395 400ctg tgg ctg tgc ttc gcc gcg atg atc agc ctg atc ggg gta ttg gct 1248Leu Trp Leu Cys Phe Ala Ala Met Ile Ser Leu Ile Gly Val Leu Ala 405 410 415tca cgc agg atg ggg gcc gac ccc gat cga tcc atg gct tga 1290Ser Arg Arg Met Gly Ala Asp Pro Asp Arg Ser Met Ala 420 425 5 429 PRT Bacterium 2412.1 5Met Thr Ala Ala Glu Glu Arg Arg Gly His Leu Gly Lys Ile Leu Arg 1 5 10 15Val Ala Ser Gly Asn Phe Leu Glu Gln Tyr Asp Phe Phe Ile Tyr Gly 20 25 30Tyr Tyr Ala Thr Tyr Ile Ala Gln Val Phe Phe Pro Ser Gly Asp Glu 35 40 45Thr Thr Ser Leu Met Leu Ser Leu Ala Thr Phe Gly Val Gly Phe Leu 50 55 60Met Arg Pro Leu Gly Ala Ile Ile Leu Gly Ser Tyr Ile Asp Arg Val 65 70 75 80Gly Arg Arg Gln Gly Leu Ile Val Thr Leu Gly Ile Met Ala Ile Gly 85 90 95Thr Leu Thr Ile Ala Leu Thr Pro Gly Tyr Ser Ala Ile Gly Ile Ala 100 105 110Ala Pro Leu Ile Val Val Ala Gly Arg Leu Leu Gln Gly Phe Ser Ala 115 120 125Gly Ala Glu Leu Gly Gly Val Ser Ile Tyr Leu Ala Glu Ile Ala Lys 130 135 140Pro Gly Arg Arg Gly Phe Tyr Thr Ser Trp Gln Ser Ala Ser Gln Gln145 150 155 160Val Ala Val Met Ala Ala Ala Leu Val Gly Leu Ser Leu Gly Ala Thr 165 170 175Leu Thr Pro Asp Gln Met His Gln Trp Gly Trp Arg Val Pro Leu Leu 180 185 190Leu Gly Cys Ala Ile Val Pro Val Ile Leu Trp Leu Arg Arg Ser Leu 195 200 205Asp Glu Thr Glu Ala Tyr Lys His Ile His His Lys Ala His Ser Leu 210 215 220Leu Gly Ser Leu Ala Gln Leu Gly Gly Ser Trp Arg Pro Ile Leu Ala225 230 235 240Gly Met Ala Ile Ser Val Leu Thr Thr Thr Thr Phe Tyr Met Ile Thr 245 250 255Ala Tyr Thr Pro Thr Phe Gly Lys Gln Ala Leu Gly Leu Asp Ala Gln 260 265 270Asp Val Leu Val Val Thr Met Leu Val Gly Ala Ser Asn Phe Ile Trp 275 280 285Leu Pro Val Gly Gly Ala Leu Ser Asp Trp Ile Gly Arg Thr Pro Val 290 295 300Leu Leu Ser Val Pro Leu Val Val Leu Val Ala Ala Tyr Pro Leu Ile305 310 315 320Ala Trp Leu Val Gly Ala Pro Ser Phe Phe Ala Phe Ala Thr Ala Leu 325 330 335Leu Ala Leu Ser Val Cys Phe Gly Leu Tyr Asn Gly Ala Met Ile Ala 340 345 350Arg Leu Thr Glu Leu Met Pro Pro Ala Ala Arg Thr Leu Gly Phe Ser 355 360 365Leu Ala Phe Ser Leu Ala Thr Ser Leu Phe Gly Gly Phe Thr Pro Leu 370 375 380Val Ser Thr Tyr Leu Ile Ser Ala Thr Gly Asn Lys Ala Ala Pro Ala385 390 395 400Leu Trp Leu Cys Phe Ala Ala Met Ile Ser Leu Ile Gly Val Leu Ala 405 410 415Ser Arg Arg Met Gly Ala Asp Pro Asp Arg Ser Met Ala 420 425 6 1677 DNA Bacterium 2412.1 fccC Flavin Monooxygenase 6atg ggg cgt gag acc cag ccg ttc gtg ctt atc gtc ggc ggc ggt caa 48Met Gly Arg Glu Thr Gln Pro Phe Val Leu Ile Val Gly Gly Gly Gln 1 5 10 15ggc ggt cta gcg ctt ggc gcg cgc ctc cgt cag ctc cag gtc ccg act 96Gly Gly Leu Ala Leu Gly Ala Arg Leu Arg Gln Leu Gln Val Pro Thr 20 25 30ctg atc gtc gat cag cac cca cgg gtg ggg gac caa tgg cga tcg cgg 144Leu Ile Val Asp Gln His Pro Arg Val Gly Asp Gln Trp Arg Ser Arg 35 40 45tac gca tcg ctc tgc ctg cac gat cca gtc tgg tac gac cac ctt cct 192Tyr Ala Ser Leu Cys Leu His Asp Pro Val Trp Tyr Asp His Leu Pro 50 55 60tac ctg ccg ttt ccc gat act tgg ccg gtt tat acg ccc aag gac aag 240Tyr Leu Pro Phe Pro Asp Thr Trp Pro Val Tyr Thr Pro Lys Asp Lys 65 70 75 80atc ggc gat tgg ctc gaa gct tat gcg cag gcg atg gag ctg ctg gtc 288Ile Gly Asp Trp Leu Glu Ala Tyr Ala Gln Ala Met Glu Leu Leu Val 85 90 95tgg tgt tcg acc aga tgc gtg tcc gcc gtc tat gac gcc gaa gcc ggg 336Trp Cys Ser Thr Arg Cys Val Ser Ala Val Tyr Asp Ala Glu Ala Gly 100 105 110cga tgg acc gtc acc ctg cgc cga ggc gag gag acc agc gtc atc cgc 384Arg Trp Thr Val Thr Leu Arg Arg Gly Glu Glu Thr Ser Val Ile Arg 115 120 125ccc gcg cat ctg gtc ctg gcg acg ggc aac gcc ggc aag ccg cgc gtt 432Pro Ala His Leu Val Leu Ala Thr Gly Asn Ala Gly Lys Pro Arg Val 130 135 140ccg cgc ttc aag ggc caa gcg cag ttc gaa ggt ccg atc ctg cac tcg 480Pro Arg Phe Lys Gly Gln Ala Gln Phe Glu Gly Pro Ile Leu His Ser145 150 155 160agc gcc tat cgg agc ggg gct gat ttc aaa gga cgg cgc gtg gcc gtg 528Ser Ala Tyr Arg Ser Gly Ala Asp Phe Lys Gly Arg Arg Val Ala Val 165 170 175atc gga tcg aac aat tcg gcc cac gac atc tgc gca gac ctc gtg gcc 576Ile Gly Ser Asn Asn Ser Ala His Asp Ile Cys Ala Asp Leu Val Ala 180 185 190cac ggc gtt gac gtc acc atg atc cag cgc agt tcg acc cat gtc gtc 624His Gly Val Asp Val Thr Met Ile Gln Arg Ser Ser Thr His Val Val 195 200 205cgt tcc gaa acg gtc atg cgg acc atg ctc gcg ccg ctt tat tca gag 672Arg Ser Glu Thr Val Met Arg Thr Met Leu Ala Pro Leu Tyr Ser Glu 210 215 220gag gcc ttg gcg gcc ggc ata ggc acg gag ctg gcc gac ctg ctt gtg 720Glu Ala Leu Ala Ala Gly Ile Gly Thr Glu Leu Ala Asp Leu Leu Val225 230 235 240gct tcc atg ccg tta cgc ctg cag gcc gaa ggc tat cgc gcc ctc cac 768Ala Ser Met Pro Leu Arg Leu Gln Ala Glu Gly Tyr Arg Ala Leu His 245 250 255gtc gcg atc gcc gag cag gac gca gcg ttc tac gcc gcg ctc gag gcg 816Val Ala Ile Ala Glu Gln Asp Ala Ala Phe Tyr Ala Ala Leu Glu Ala 260 265 270atc ggc ttc atg cat gac ttc ggc gag gac ggc acc ggc atg ccg ctg 864Ile Gly Phe Met His Asp Phe Gly Glu Asp Gly Thr Gly Met Pro Leu 275 280 285aag tat ctt cgt cgc gcg tcg ggg tac tat atc gac gtc ggc gca tcc 912Lys Tyr Leu Arg Arg Ala Ser Gly Tyr Tyr Ile Asp Val Gly Ala Ser 290 295 300gaa ctc ctg gcc agc ggg gcc ata aag ctg cgc tcc cgc gtc gag atc 960Glu Leu Leu Ala Ser Gly Ala Ile Lys Leu Arg Ser Arg Val Glu Ile305 310 315 320gat cac ttc gac acc gac ggc ctg gcc ctc tcg gac ggc agc aag gtc 1008Asp His Phe Asp Thr Asp Gly Leu Ala Leu Ser Asp Gly Ser Lys Val 325 330 335gac gcc gac gcc gtc atc tgc gca acc ggt ttc ggc tcc atg gac gag 1056Asp Ala Asp Ala Val Ile Cys Ala Thr Gly Phe Gly Ser Met Asp Glu 340 345 350tgg gcg gcc gaa ttg att tcc ccc gag gtc gcg gcc aag gtc gga agg 1104Trp Ala Ala Glu Leu Ile Ser Pro Glu Val Ala Ala Lys Val Gly Arg 355 360 365gtc tgg ggc tat ggg tcc ggc acc cga ggc gat ccg ggc ccc tgg gag 1152Val Trp Gly Tyr Gly Ser Gly Thr Arg Gly Asp Pro Gly Pro Trp Glu 370 375 380ggc gaa ctt cgg aac atg tgg aag ccc acc cgc cag cag ggc ttg tgg 1200Gly Glu Leu Arg Asn Met Trp Lys Pro Thr Arg Gln Gln Gly Leu Trp385 390 395 400ttc cag ggc gga aac ctg gcg caa acc cgc ttc tac tcc aga gcg ctc 1248Phe Gln Gly Gly Asn Leu Ala Gln Thr Arg Phe Tyr Ser Arg Ala Leu 405 410 415gct ctg cag ttg aag ccc gac atg ctg att gcc gtg agt cta cgt tcg 1296Ala Leu Gln Leu Lys Pro Asp Met Leu Ile Ala Val Ser Leu Arg Ser 420 425 430tca ccg act agg cgg cgg cga agc tca ctg atc ttg ctc cgc ggc gtc 1344Ser Pro Thr Arg Arg Arg Arg Ser Ser Leu Ile Leu Leu Arg Gly Val 435 440 445cca gag gct cac gct cgc ctc tcc ctg gtc tat tgc ttc gcg aat ttg 1392Pro Glu Ala His Ala Arg Leu Ser Leu Val Tyr Cys Phe Ala Asn Leu 450 455 460cgg ccg cag ctc gag atg ttc ggg caa gaa cgt cct gca ata ggc gac 1440Arg Pro Gln Leu Glu Met Phe Gly Gln Glu Arg Pro Ala Ile Gly Asp465 470 475 480gct cac ggt cag ggc ctc ttc gta gta gtc ctc ggt gat cgc acc gtt 1488Ala His Gly Gln Gly Leu Phe Val Val Val Leu Gly Asp Arg Thr Val 485 490 495ttg gac aaa cga gat cgc cag aac cgc atc gac gat ctg gat gtt cgt 1536Leu Asp Lys Arg Asp Arg Gln Asn Arg Ile Asp Asp Leu Asp Val Arg 500 505 510gtg aaa ctt ctt ctg cgg atc gcg cag aaa agg cat gtg gaa gat gcg 1584Val Lys Leu Leu Leu Arg Ile Ala Gln Lys Arg His Val Glu Asp Ala 515 520 525gtc aag gcg ctg ata ggc cgc gcg agc gac ggc ttc cac ata ttc gcg 1632Val Lys Ala Leu Ile Gly Arg Ala Ser Asp Gly Phe His Ile Phe Ala 530 535 540atc ggc ctg ccg tgt ctc gag ccc gcc aaa tcc acc cag gaa tag 1677Ile Gly Leu Pro Cys Leu Glu Pro Ala Lys Ser Thr Gln Glu545 550 555 7 558 PRT Bacterium 2412.1 7Met Gly Arg Glu Thr Gln Pro Phe Val Leu Ile Val Gly Gly Gly Gln 1 5 10 15Gly Gly Leu Ala Leu Gly Ala Arg Leu Arg Gln Leu Gln Val Pro Thr 20 25 30Leu Ile Val Asp Gln His Pro Arg Val Gly Asp Gln Trp Arg Ser Arg 35 40 45Tyr Ala Ser Leu Cys Leu His Asp Pro Val Trp Tyr Asp His Leu Pro 50 55 60Tyr Leu Pro Phe Pro Asp Thr Trp Pro Val Tyr Thr Pro Lys Asp Lys 65 70 75 80Ile Gly Asp Trp Leu Glu Ala Tyr Ala Gln Ala Met Glu Leu Leu Val 85 90 95Trp Cys Ser Thr Arg Cys Val Ser Ala Val Tyr Asp Ala Glu Ala Gly 100 105 110Arg Trp Thr Val Thr Leu Arg Arg Gly Glu Glu Thr Ser Val Ile Arg 115 120 125Pro Ala His Leu Val Leu Ala Thr Gly Asn Ala Gly Lys Pro Arg Val 130 135 140Pro Arg Phe Lys Gly Gln Ala Gln Phe Glu Gly Pro Ile Leu His Ser145 150 155 160Ser Ala Tyr Arg Ser Gly Ala Asp Phe Lys Gly Arg Arg Val Ala Val 165 170 175Ile Gly Ser Asn Asn Ser Ala His Asp Ile Cys Ala Asp Leu Val Ala 180 185 190His Gly Val Asp Val Thr Met Ile Gln Arg Ser Ser Thr His Val Val 195 200 205Arg Ser Glu Thr Val Met Arg Thr Met Leu Ala Pro Leu Tyr Ser Glu 210 215 220Glu Ala Leu Ala Ala Gly Ile Gly Thr Glu Leu Ala Asp Leu Leu Val225 230 235 240Ala Ser Met Pro Leu Arg Leu Gln Ala Glu Gly Tyr Arg Ala Leu His 245 250 255Val Ala Ile Ala Glu Gln Asp Ala Ala Phe Tyr Ala Ala Leu Glu Ala 260 265 270Ile Gly Phe Met His Asp Phe Gly Glu Asp Gly Thr Gly Met Pro Leu 275 280 285Lys Tyr Leu Arg Arg Ala Ser Gly Tyr Tyr Ile Asp Val Gly Ala Ser 290 295 300Glu Leu Leu Ala Ser Gly Ala Ile Lys Leu Arg Ser Arg Val Glu Ile305 310 315 320Asp His Phe Asp Thr Asp Gly Leu Ala Leu Ser Asp Gly Ser Lys Val 325 330 335Asp Ala Asp Ala Val Ile Cys Ala Thr Gly Phe Gly Ser Met Asp Glu 340 345 350Trp Ala Ala Glu Leu Ile Ser Pro Glu Val Ala Ala Lys Val Gly Arg 355 360 365Val Trp Gly Tyr Gly Ser Gly Thr Arg Gly Asp Pro Gly Pro Trp Glu 370 375 380Gly Glu Leu Arg Asn Met Trp Lys Pro Thr Arg Gln Gln Gly Leu Trp385 390 395 400Phe Gln Gly Gly Asn Leu Ala Gln Thr Arg Phe Tyr Ser Arg Ala Leu 405 410 415Ala Leu Gln Leu Lys Pro Asp Met Leu Ile Ala Val Ser Leu Arg Ser 420 425 430Ser Pro Thr Arg Arg Arg Arg Ser Ser Leu Ile Leu Leu Arg Gly Val 435 440 445Pro Glu Ala His Ala Arg Leu Ser Leu Val Tyr Cys Phe Ala Asn Leu 450 455 460Arg Pro Gln Leu Glu Met Phe Gly Gln Glu Arg Pro Ala Ile Gly Asp465 470 475 480Ala His Gly Gln Gly Leu Phe Val Val Val Leu Gly Asp Arg Thr Val 485 490 495Leu Asp Lys Arg Asp Arg Gln Asn Arg Ile Asp Asp Leu Asp Val Arg 500 505 510Val Lys Leu Leu Leu Arg Ile Ala Gln Lys Arg His Val Glu Asp Ala 515 520 525Val Lys Ala Leu Ile Gly Arg Ala Ser Asp Gly Phe His Ile Phe Ala 530 535 540Ile Gly Leu Pro Cys Leu Glu Pro Ala Lys Ser Thr Gln Glu545 550 555 8 1407 DNA Bacterium 2412.1 fccD Aldehyde Dehydrogenase 8atg cta gag tac aag ctg ctg atc gac ggc cgc ctg gtc gcc ggc gca 48Met Leu Glu Tyr Lys Leu Leu Ile Asp Gly Arg Leu Val Ala Gly Ala 1 5 10 15acg acg atg tcc gta atc aat ccc gcg act gaa acg ccg ctg gtg atc 96Thr Thr Met Ser Val Ile Asn Pro Ala Thr Glu Thr Pro Leu Val Ile 20 25 30gat tgc ccc agg gcc gac cgc gac caa ctg gac gaa gcc gtc gcc gcc 144Asp Cys Pro Arg Ala Asp Arg Asp Gln Leu Asp Glu Ala Val Ala Ala 35 40 45gcg gaa cgc gcc ttt cag agc tgg cgg gca acc acg ctc gag cag cgc 192Ala Glu Arg Ala Phe Gln Ser Trp Arg Ala Thr Thr Leu Glu Gln Arg 50 55 60agg gcc acg ctc aac gcc atc gcc gac gca att gaa gcc gac cag tcg 240Arg Ala Thr Leu Asn Ala Ile Ala Asp Ala Ile Glu Ala Asp Gln Ser 65 70 75 80gcg ctg gcg cgt ttg ctg acg cag gaa cag ggc aag ccg ctc gca gac 288Ala Leu Ala Arg Leu Leu Thr Gln Glu Gln Gly Lys Pro Leu Ala Asp 85 90 95gcg atg ggc gag atc tac gcc tcc gcg gcc ttc ttc cgc tac ttc acc 336Ala Met Gly Glu Ile Tyr Ala Ser Ala Ala Phe Phe Arg Tyr Phe Thr 100 105 110tcg ctc gat ctg ccg cct cgc gtg gtc aga gac gac gcg acg ggc cgc 384Ser Leu Asp Leu Pro Pro Arg Val Val Arg Asp Asp Ala Thr Gly Arg 115 120 125gta gag gtg cat aga cgc ccc cta ggc gtg gtg ggc tgc atc gtc ccc 432Val Glu Val His Arg Arg Pro Leu Gly Val Val Gly Cys Ile Val Pro 130 135 140tgg aat ttc ccg atg ctg ttg atg gcg ttc aag atc ccg gcg gcc ctg 480Trp Asn Phe Pro Met Leu Leu Met Ala Phe Lys Ile Pro Ala Ala Leu145 150 155 160ctg gcc ggc aac acg gtc atc ctc aag ccg gcg gcg acg acg cct ctg 528Leu Ala Gly Asn Thr Val Ile Leu Lys Pro Ala Ala Thr Thr Pro Leu 165 170 175acg gcg ctt cgg ttt ggc gcc ttg gtc aag gat atc gtc cca ccg ggc 576Thr Ala Leu Arg Phe Gly Ala Leu Val Lys Asp Ile Val Pro Pro Gly 180 185 190gtc att aac atc atc acc gac gcc gac gat ctc ggc gcg gaa atg acc 624Val Ile Asn Ile Ile Thr Asp Ala Asp Asp Leu Gly Ala Glu Met Thr 195 200 205cgc cat cct ggc att cgc aaa atc agc ttc acc gga tcg acc cag acc 672Arg His Pro Gly Ile Arg Lys Ile Ser Phe Thr Gly Ser Thr Gln Thr 210 215 220gga aaa aag gtc atg gcc ggc gcg gcc gaa ggc ctc aaa cgt ata tcg 720Gly Lys Lys Val Met Ala Gly Ala Ala Glu Gly Leu Lys Arg Ile Ser225 230 235 240ctt gag ttg ggc gga aac gac gct ctg atc gtc ctg gat gac gtc gac 768Leu Glu Leu Gly Gly Asn Asp Ala Leu Ile Val Leu Asp Asp Val Asp 245 250 255ccc aag gaa gtc gct ccc agg gtg ttc gcc gcg gcc atg caa aac gcc 816Pro Lys Glu Val Ala Pro Arg Val Phe Ala Ala Ala Met Gln Asn Ala 260 265 270ggt cag gtg tgc atc gcc gcc aaa cgg att tac gtc cat gag agc ctc 864Gly Gln Val Cys Ile Ala Ala Lys Arg Ile Tyr Val His Glu Ser Leu 275 280 285tat gag gcc atg tgc gag gag ttc gcg cag ttg gcg gcc cgc acg gtc 912Tyr Glu Ala Met Cys Glu Glu Phe Ala Gln Leu Ala Ala Arg Thr Val 290 295 300gtg ggc gat gga ctc gaa cag ggc gtt cag atg ggg ccg ctg cag aac 960Val Gly Asp Gly Leu Glu Gln Gly Val Gln Met Gly Pro Leu Gln Asn305 310 315 320cgg cgc cag ttc gag aag gtt ctt ggt cta atc gag cgt gcg agg acg 1008Arg Arg Gln Phe Glu Lys Val Leu Gly Leu Ile Glu Arg Ala Arg Thr 325 330 335gac ggc cgc atc atc gcc ggc ggc cgc cgc aag ggc gac aag ggc tat 1056Asp Gly Arg Ile Ile Ala Gly Gly Arg Arg Lys Gly Asp Lys Gly Tyr 340 345 350ttc atc gag ccc acc atc gta cgc gac atc gcc gaa ggc gct cag ctc 1104Phe Ile Glu Pro Thr Ile Val Arg Asp Ile Ala Glu Gly Ala Gln Leu 355 360 365gtc gac gaa gag cag ttt ggc ccg gtg atg ccg gtg atc cgg tac tcc 1152Val Asp Glu Glu Gln Phe Gly Pro Val Met Pro Val Ile Arg Tyr Ser 370 375 380gac ccc gtc gac gcc gtg cgc cgc gcc aac gcc tcg ccc tat ggt ctg 1200Asp Pro Val Asp Ala Val Arg Arg Ala Asn Ala Ser Pro Tyr Gly Leu385 390 395 400ggg gga tcc atc tgg tcc cgc aac gtc gtc cgc gcg tgg agc ctg gcc 1248Gly Gly Ser Ile Trp Ser Arg Asn Val Val Arg Ala Trp Ser Leu Ala 405 410 415gcc gat atg gag gcc ggc tcg gtc tgg gtg aac aag cac gcc gac gtg 1296Ala Asp Met Glu Ala Gly Ser Val Trp Val Asn Lys His Ala Asp Val 420 425 430cag ccc gat ctc ccg ctc ggc ggc gcc aag ttc tcg ggg atg ggc tcg 1344Gln Pro Asp Leu Pro Leu Gly Gly Ala Lys Phe Ser Gly Met Gly Ser 435 440 445gag tta ggc gag gaa ggg ctg cac gag ttc acc caa gtg cag gtg ctg 1392Glu Leu Gly Glu Glu Gly Leu His Glu Phe Thr Gln Val Gln Val Leu 450 455 460aat atg acg cgg tga 1407Asn Met Thr Arg465 9 468 PRT Bacterium 2412.1 9Met Leu Glu Tyr Lys Leu Leu Ile Asp Gly Arg Leu Val Ala Gly Ala 1 5 10 15Thr Thr Met Ser Val Ile Asn Pro Ala Thr Glu Thr Pro Leu Val Ile 20 25 30Asp Cys Pro Arg Ala Asp Arg Asp Gln Leu Asp Glu Ala Val Ala Ala 35 40 45Ala Glu Arg Ala Phe Gln Ser Trp Arg Ala Thr Thr Leu Glu Gln Arg 50 55 60Arg Ala Thr Leu Asn Ala Ile Ala Asp Ala Ile Glu Ala Asp Gln Ser 65 70 75 80Ala Leu Ala Arg Leu Leu Thr Gln Glu Gln Gly Lys Pro Leu Ala Asp 85 90 95Ala Met Gly Glu Ile Tyr Ala Ser Ala Ala Phe Phe Arg Tyr Phe Thr 100 105 110Ser Leu Asp Leu Pro Pro Arg Val Val Arg Asp Asp Ala Thr Gly Arg 115 120 125Val Glu Val His Arg Arg Pro Leu Gly Val Val Gly Cys Ile Val Pro 130 135 140Trp Asn Phe Pro Met Leu Leu Met Ala Phe Lys Ile Pro Ala Ala Leu145 150 155 160Leu Ala Gly Asn Thr Val Ile Leu Lys Pro Ala Ala Thr Thr Pro Leu 165 170 175Thr Ala Leu Arg Phe Gly Ala Leu Val Lys Asp Ile Val Pro Pro Gly 180 185 190Val Ile Asn Ile Ile Thr Asp Ala Asp Asp Leu Gly Ala Glu Met Thr 195 200 205Arg His Pro Gly Ile Arg Lys Ile Ser Phe Thr Gly Ser Thr Gln Thr 210 215 220Gly Lys Lys Val Met Ala Gly Ala Ala Glu Gly Leu Lys Arg Ile Ser225 230 235 240Leu Glu Leu Gly Gly Asn Asp Ala Leu Ile Val Leu Asp Asp Val Asp 245 250 255Pro Lys Glu Val Ala Pro Arg Val Phe Ala Ala Ala Met Gln Asn Ala 260 265 270Gly Gln Val Cys Ile Ala Ala Lys Arg Ile Tyr Val His Glu Ser Leu 275 280 285Tyr Glu Ala Met Cys Glu Glu Phe Ala Gln Leu Ala Ala Arg Thr Val 290 295 300Val Gly Asp Gly Leu Glu Gln Gly Val Gln Met Gly Pro Leu Gln Asn305 310 315 320Arg Arg Gln Phe Glu Lys Val Leu Gly Leu Ile Glu Arg Ala Arg Thr 325 330 335Asp Gly Arg Ile Ile Ala Gly Gly Arg Arg Lys Gly Asp Lys Gly Tyr 340 345 350Phe Ile Glu Pro Thr Ile Val Arg Asp Ile Ala Glu Gly Ala Gln Leu 355 360 365Val Asp Glu Glu Gln Phe Gly Pro Val Met Pro Val Ile Arg Tyr Ser 370 375 380Asp Pro Val Asp Ala Val Arg Arg Ala Asn Ala Ser Pro Tyr Gly Leu385 390 395 400Gly Gly Ser Ile Trp Ser Arg Asn Val Val Arg Ala Trp Ser Leu Ala 405 410 415Ala Asp Met Glu Ala Gly Ser Val Trp Val Asn Lys His Ala Asp Val 420 425 430Gln Pro Asp Leu Pro Leu Gly Gly Ala Lys Phe Ser Gly Met Gly Ser 435 440 445Glu Leu Gly Glu Glu Gly Leu His Glu Phe Thr Gln Val Gln Val Leu 450 455 460Asn Met Thr Arg465 10 939 DNA Bacterium 2412.1 fccE Alcohol Dehydrogenase 10atg aaa gcg gcc att tac cgg cgc ggg gag atc gtt gtc gat acc gtt 48Met Lys Ala Ala Ile Tyr Arg Arg Gly Glu Ile Val Val Asp Thr Val 1 5 10 15ccc gat ccg gtt cca gga cca ggc cag gtt ctc gtc cgg agc ctt gtt 96Pro Asp Pro Val Pro Gly Pro Gly Gln Val Leu Val Arg Ser Leu Val 20 25 30tgc ggg gta tgt ggt tcg gat ctg cat tac cga cat cac gca cac cgg 144Cys Gly Val Cys Gly Ser Asp Leu His Tyr Arg His His Ala His Arg 35 40 45ttc gtc gat ctg gcc ttg cgc tcg ggc gcg ccc gcc ctg gcc gcc gat 192Phe Val Asp Leu Ala Leu Arg Ser Gly Ala Pro Ala Leu Ala Ala Asp 50 55 60ttg gat cgc gat atc gtc ctt ggt cac gaa ttc agc gct caa gtc gtc 240Leu Asp Arg Asp Ile Val Leu Gly His Glu Phe Ser Ala Gln Val Val 65 70 75 80gac tac ggg cct aag acc gag cgt ctc ctg aag tcg gga acg gtc gtc 288Asp Tyr Gly Pro Lys Thr Glu Arg Leu Leu Lys Ser Gly Thr Val Val 85 90 95tgc tcg ccc ccc gtc gcg ttc ggg gcc agc ggc atg cgc gcc gtt ggc 336Cys Ser Pro Pro Val Ala Phe Gly Ala Ser Gly Met Arg Ala Val Gly 100 105 110tac tcc gac gaa tta ccg ggc ggg ttt ggc cag tac atg gtc ttg aat 384Tyr Ser Asp Glu Leu Pro Gly Gly Phe Gly Gln Tyr Met Val Leu Asn 115 120 125gag gcg ttc ctg atg ccg gcc cca aac gga ctg gat ccg gct cgc gcg 432Glu Ala Phe Leu Met Pro Ala Pro Asn Gly Leu Asp Pro Ala Arg Ala 130 135 140gcg ctc acc gag ccg atg gcg gtg ggg tgg cac gcg gtg aag ctg gcc 480Ala Leu Thr Glu Pro Met Ala Val Gly Trp His Ala Val Lys Leu Ala145 150 155 160ggt ccc gga cgc gac cat atc ccg ctc gtg atc ggc tgc ggg ccc gtg 528Gly Pro Gly Arg Asp His Ile Pro Leu Val Ile Gly Cys Gly Pro Val 165 170 175ggc atg gcg gtc atc gcc gcg ctc cgg ggt ctg ggc gtc gga ccg atc 576Gly Met Ala Val Ile Ala Ala Leu Arg Gly Leu Gly Val Gly Pro Ile 180 185 190atc gcg gcc gac ttc aat ccg gcg cgt cgg agc ctg gcg gcg cgc atg 624Ile Ala Ala Asp Phe Asn Pro Ala Arg Arg Ser Leu Ala Ala Arg Met 195 200 205ggc gcc gat att gtc atc gac ccg gcg gag cgg tcc ccc tac gac gaa 672Gly Ala Asp Ile Val Ile Asp Pro Ala Glu Arg Ser Pro Tyr Asp Glu 210 215 220tgg cgg gat acc gcg gcg gcg tca ggc ctg gcc gga ctg gcg ggg gcg 720Trp Arg Asp Thr Ala Ala Ala Ser Gly Leu Ala Gly Leu Ala Gly Ala225 230 235 240cca gcg tcg ctg cgg acc tgt ctg gtc ttc gag tgt gtc ggc ctg cca 768Pro Ala Ser Leu Arg Thr Cys Leu Val Phe Glu Cys Val Gly Leu Pro 245 250 255gga atg ctg cgt cag atc atg gaa ggc gcc ccg gcg gag tcg gag atc 816Gly Met Leu Arg Gln Ile Met Glu Gly Ala Pro Ala Glu Ser Glu Ile 260 265 270atc gtc gtc ggg gcc tgc atg gag ccc gat agc ctc gag ccg atg atg 864Ile Val Val Gly Ala Cys Met Glu Pro Asp Ser Leu Glu Pro Met Met 275 280 285gcg atg cat aag gct ctg acg ctg aat ttt cgc gaa cct aca cga tcg 912Ala Met His Lys Ala Leu Thr Leu Asn Phe Arg Glu Pro Thr Arg Ser 290 295 300agg agt tcg ccg agg tcc ttc gga tga 939Arg Ser Ser Pro Arg Ser Phe Gly305 310 11 312 PRT Bacterium 2412.1 11Met Lys Ala Ala Ile Tyr Arg Arg Gly Glu Ile Val Val Asp Thr Val 1 5 10 15Pro Asp Pro Val Pro Gly Pro Gly Gln Val Leu Val Arg Ser Leu Val 20 25 30Cys Gly Val Cys Gly Ser Asp Leu His Tyr Arg His His Ala His Arg 35 40 45Phe Val Asp Leu Ala Leu Arg Ser Gly Ala Pro Ala Leu Ala Ala Asp 50 55 60Leu Asp Arg Asp Ile Val Leu Gly His Glu Phe Ser Ala Gln Val Val 65 70 75 80Asp Tyr Gly Pro Lys Thr Glu Arg Leu Leu Lys Ser Gly Thr Val Val 85 90 95Cys Ser Pro Pro Val Ala Phe Gly Ala Ser Gly Met Arg Ala Val Gly 100 105 110Tyr Ser Asp Glu Leu Pro Gly Gly Phe Gly Gln Tyr Met Val Leu Asn 115 120 125Glu Ala Phe Leu Met Pro Ala Pro Asn Gly Leu Asp Pro Ala Arg Ala 130 135 140Ala Leu Thr Glu Pro Met Ala Val Gly Trp His Ala Val Lys Leu Ala145 150 155 160Gly Pro Gly Arg Asp His Ile Pro Leu Val Ile Gly Cys Gly Pro Val 165 170 175Gly Met Ala Val Ile Ala Ala Leu Arg Gly Leu Gly Val Gly Pro Ile 180 185 190Ile Ala Ala Asp Phe Asn Pro Ala Arg Arg Ser Leu Ala Ala Arg Met 195 200 205Gly Ala Asp Ile Val Ile Asp Pro Ala Glu Arg Ser Pro Tyr Asp Glu 210 215 220Trp Arg Asp Thr Ala Ala Ala Ser Gly Leu Ala Gly Leu Ala Gly Ala225 230 235 240Pro Ala Ser Leu Arg Thr Cys Leu Val Phe Glu Cys Val Gly Leu Pro 245 250 255Gly Met Leu Arg Gln Ile Met Glu Gly Ala Pro Ala Glu Ser Glu Ile 260 265 270Ile Val Val Gly Ala Cys Met Glu Pro Asp Ser Leu Glu Pro Met Met 275 280 285Ala Met His Lys Ala Leu Thr Leu Asn Phe Arg Glu Pro Thr Arg Ser 290 295 300Arg Ser Ser Pro Arg Ser Phe Gly305 310 12 1377 DNA Bacterium 2412.1 fccF CoA Ligase 12atg ctt ggc gcc gtc atc cag acg gtg aac atc cga cta gcc cga gac 48Met Leu Gly Ala Val Ile Gln Thr Val Asn Ile Arg Leu Ala Arg Asp 1 5 10 15gac ctg cgc tac acg ctc gag cat gcg ggc gcc acc ctg gcg ctg agc 96Asp Leu Arg Tyr Thr Leu Glu His Ala Gly Ala Thr Leu Ala Leu Ser 20 25 30cac acc gat ttc ctg ccg atc ctc gag gag gtg atc gac caa ttg ccc 144His Thr Asp Phe Leu Pro Ile Leu Glu Glu Val Ile Asp Gln Leu Pro 35 40 45agc ctg cgc ggg gtc gtc cat ctg aag gac gac gag gcg gaa gcc gcc 192Ser Leu Arg Gly Val Val His Leu Lys Asp Asp Glu Ala Glu Ala Ala 50 55 60cat ccc tgg gtg ctg ggg gag tat gag gcc ctg atg gcg gcc gcg cgc 240His Pro Trp Val Leu Gly Glu Tyr Glu Ala Leu Met Ala Ala Ala Arg 65 70 75 80cct cgg ttc gac ttc ccg gac ttc gac gag aac acg cgg gcg acg acc 288Pro Arg Phe Asp Phe Pro Asp Phe Asp Glu Asn Thr Arg Ala Thr Thr 85 90 95ttc tac acc agc ggc acg acc ggg cgt ccg aag ggc gtc tac tat tcg 336Phe Tyr Thr Ser Gly Thr Thr Gly Arg Pro Lys Gly Val Tyr Tyr Ser 100 105 110cat cgt cag ctg gtg ctg cac acc ctg gcg gtg atg gcg acg ctg gcc 384His Arg Gln Leu Val Leu His Thr Leu Ala Val Met Ala Thr Leu Ala 115 120 125ctt gga gac ggt tac gcc agg ctg ggg cgc gat acg gtc tac atg ccg 432Leu Gly Asp Gly Tyr Ala Arg Leu Gly Arg Asp Thr Val Tyr Met Pro 130 135 140atc acc ccg atg ttc cat gct cat gcg tgg gga atg ccc ttc gtg gcg 480Ile Thr Pro Met Phe His Ala His Ala Trp Gly Met Pro Phe Val Ala145 150 155 160acg atg gtc ggc tgc aag caa gtc tac cca ggg cgc tat gtt ccc gag 528Thr Met Val Gly Cys Lys Gln Val Tyr Pro Gly Arg Tyr Val Pro Glu 165 170 175caa ctg gtg gag ctt cag cgc gcg gag aag gtg acc ttc tct cat tgc 576Gln Leu Val Glu Leu Gln Arg Ala Glu Lys Val Thr Phe Ser His Cys 180 185 190gtg ccc aca ctt ttg cag atg atg ctc aat tcg cct tcg ggc cag acg 624Val Pro Thr Leu Leu Gln Met Met Leu Asn Ser Pro Ser Gly Gln Thr 195 200 205gcg gat ttc acc gga tgg cag gtg ctc gtc ggc gga gcg gcg ctg ccc 672Ala Asp Phe Thr Gly Trp Gln Val Leu Val Gly Gly Ala Ala Leu Pro 210 215 220cgc ggc ctg gct ctt cag gcc gcg ggg cgc ggc atc gtc ctg acc acc 720Arg Gly Leu Ala Leu Gln Ala Ala Gly Arg Gly Ile Val Leu Thr Thr225 230 235 240gga tac gga atg tcc gaa acc ggg ccg ctg gtc agc ttc acg cgc att 768Gly Tyr Gly Met Ser Glu Thr Gly Pro Leu Val Ser Phe Thr Arg Ile 245 250 255agg acc gaa gca atg gct cca gct cag gag gag gtc gcc att cgc acc 816Arg Thr Glu Ala Met Ala Pro Ala Gln Glu Glu Val Ala Ile Arg Thr 260 265 270aag gtc gga caa gct atc gcg ctg gtc gac ctc cgg gtc gtg gat gag 864Lys Val Gly Gln Ala Ile Ala Leu Val Asp Leu Arg Val Val Asp Glu 275 280 285tcc atg gcg gat gtg ccc cgc gac ggc ctc tcc gcg ggc gag atc gtg 912Ser Met Ala Asp Val Pro Arg Asp Gly Leu Ser Ala Gly Glu Ile Val 290 295 300ttg cgt gcg cct tgg ctg acg gct ggg tac cat cgc gat ctg gcc gcc 960Leu Arg Ala Pro Trp Leu Thr Ala Gly Tyr His Arg Asp Leu Ala Ala305 310 315 320tcg cgc gag ctt tgg cgc gga gga agc ctt cat acg cag gat ttc ggc 1008Ser Arg Glu Leu Trp Arg Gly Gly Ser Leu His Thr Gln Asp Phe Gly 325 330 335cgg att gac gcg gag ggc tac ctg cag atc agc gac cgc ctc cag gga 1056Arg Ile Asp Ala Glu Gly Tyr Leu Gln Ile Ser Asp Arg Leu Gln Gly 340 345 350gtc atc aag acg gtg ggg atg ggt tct cct gag ctg gga gat ctc gtc 1104Val Ile Lys Thr Val Gly Met Gly Ser Pro Glu Leu Gly Asp Leu Val 355 360 365agc cgc cat ccg gcg gtg ctg gag agc gcc gcg atc gct gtc gcc gac 1152Ser Arg His Pro Ala Val Leu Glu Ser Ala Ala Ile Ala Val Ala Asp 370 375 380gag cgt tgg gga gag cgc cca gcg atg gtc gtc gtg ctc agg ccg ggc 1200Glu Arg Trp Gly Glu Arg Pro Ala Met Val Val Val Leu Arg Pro Gly385 390 395 400atg agc gcg acc acg gcg gac atc cga gac cac ctt tca tcg tat gtc 1248Met Ser Ala Thr Thr Ala Asp Ile Arg Asp His Leu Ser Ser Tyr Val 405 410 415gcg acc ggc gaa ata cct cgc tac gcc gtg ccc gag cag atc tgg ttc 1296Ala Thr Gly Glu Ile Pro Arg Tyr Ala Val Pro Glu Gln Ile Trp Phe 420 425 430gtc gag gag ctc gac cga acg agc gtg ggc aag gtc gac aag cgg gcg 1344Val Glu Glu Leu Asp Arg Thr Ser Val Gly Lys Val Asp Lys Arg Ala 435 440 445ctt cgt tcc agg ttc gcc gaa gcg gcg tcc tga 1377Leu Arg Ser Arg Phe Ala Glu Ala Ala Ser 450 455 13 458 PRT Bacterium 2412.1 13Met Leu Gly Ala Val Ile Gln Thr Val Asn Ile Arg Leu Ala Arg Asp 1 5 10 15Asp Leu Arg Tyr Thr Leu Glu His Ala Gly Ala Thr Leu Ala Leu Ser 20 25 30His Thr Asp Phe Leu Pro Ile Leu Glu Glu Val Ile Asp Gln Leu Pro 35 40 45Ser Leu Arg Gly Val Val His Leu Lys Asp Asp Glu Ala Glu Ala Ala 50 55 60His Pro Trp Val Leu Gly Glu Tyr Glu Ala Leu Met Ala Ala Ala Arg 65 70 75 80Pro Arg Phe Asp Phe Pro Asp Phe Asp Glu Asn Thr Arg Ala Thr Thr 85 90 95Phe Tyr Thr Ser Gly Thr Thr Gly Arg Pro Lys Gly Val Tyr Tyr Ser 100 105 110His Arg Gln Leu Val Leu His Thr Leu Ala Val Met Ala Thr Leu Ala 115 120 125Leu Gly Asp Gly Tyr Ala Arg Leu Gly Arg Asp Thr Val Tyr Met Pro 130 135 140Ile Thr Pro Met Phe His Ala His Ala Trp Gly Met Pro Phe Val Ala145 150 155 160Thr Met Val Gly Cys Lys Gln Val Tyr Pro Gly Arg Tyr Val Pro Glu 165 170 175Gln Leu Val Glu Leu Gln Arg Ala Glu Lys Val Thr Phe Ser His Cys 180 185 190Val Pro Thr Leu Leu Gln Met Met Leu Asn Ser Pro Ser Gly Gln Thr 195 200 205Ala Asp Phe Thr Gly Trp Gln Val Leu Val Gly Gly Ala Ala Leu Pro 210 215 220Arg Gly Leu Ala Leu Gln Ala Ala Gly Arg Gly Ile Val Leu Thr Thr225 230 235 240Gly Tyr Gly Met Ser Glu Thr Gly Pro Leu Val Ser Phe Thr Arg Ile 245 250 255Arg Thr Glu Ala Met Ala Pro Ala Gln Glu Glu Val Ala Ile Arg Thr 260 265 270Lys Val Gly Gln Ala Ile Ala Leu Val Asp Leu Arg Val Val Asp Glu 275 280 285Ser Met Ala Asp Val Pro Arg Asp Gly Leu Ser Ala Gly Glu Ile Val 290 295 300Leu Arg Ala Pro Trp Leu Thr Ala Gly Tyr His Arg Asp Leu Ala Ala305 310 315 320Ser Arg Glu Leu Trp Arg Gly Gly Ser Leu His Thr Gln Asp Phe Gly 325 330 335Arg Ile Asp Ala Glu Gly Tyr Leu Gln Ile Ser Asp Arg Leu Gln Gly 340 345 350Val Ile Lys Thr Val Gly Met Gly Ser Pro Glu Leu Gly Asp Leu Val 355 360 365Ser Arg His Pro Ala Val Leu Glu Ser Ala Ala Ile Ala Val Ala Asp 370 375 380Glu Arg Trp Gly Glu Arg Pro Ala Met Val Val Val Leu Arg Pro Gly385 390 395 400Met Ser Ala Thr Thr Ala Asp Ile Arg Asp His Leu Ser Ser Tyr Val 405 410 415Ala Thr Gly Glu Ile Pro Arg Tyr Ala Val Pro Glu Gln Ile Trp Phe 420 425 430Val Glu Glu Leu Asp Arg Thr Ser Val Gly Lys Val Asp Lys Arg Ala 435 440 445Leu Arg Ser Arg Phe Ala Glu Ala Ala Ser 450 455 14 1632 DNA Bacterium 2412.1 fccG Acetohydroxyacid synthase 14atg tcg ccg gag gtt gat ccg ctg ctg gcg gcg ctc gac gac aac ggc 48Met Ser Pro Glu Val Asp Pro Leu Leu Ala Ala Leu Asp Asp Asn Gly 1 5 10 15atc cgc ttc atc ccc gtg cgg cat gag gcg gct gcg gcc tat atg gcc 96Ile Arg Phe Ile Pro Val Arg His Glu Ala Ala Ala Ala Tyr Met Ala 20 25 30gag ggt ctc tac aag acg aca ggc caa gtc gcc gcc acc gtg aca aac 144Glu Gly Leu Tyr Lys Thr Thr Gly Gln Val Ala Ala Thr Val Thr Asn 35 40 45cca gga ccc ggt acg gca aac ctg ctg ccc ggc ttg gtg acc gcc aag 192Pro Gly Pro Gly Thr Ala Asn Leu Leu Pro Gly Leu Val Thr Ala Lys 50 55 60cat gag ggc gtg ccg atg atc gcc atc acc gcg cag cat cat ggc ggg 240His Glu Gly Val Pro Met Ile Ala Ile Thr Ala Gln His His Gly Gly 65 70 75 80gtc gtc tat ccc gcg acg ccc agc acg ttc cag ggc gcc gac caa ttg 288Val Val Tyr Pro Ala Thr Pro Ser Thr Phe Gln Gly Ala Asp Gln Leu 85 90 95gaa ctc ctg cgc ccg gcg gtc aaa tgg ggc gcg ccc att cat acc tgg 336Glu Leu Leu Arg Pro Ala Val Lys Trp Gly Ala Pro Ile His Thr Trp 100 105 110caa cgc atc ggc gag gtt acg cgc atg gcg ttc cgg gag atg tgg gcg 384Gln Arg Ile Gly Glu Val Thr Arg Met Ala Phe Arg Glu Met Trp Ala 115 120 125ggg cgt ccc ggc ccg gtc cag atc gat gtt ccg agc cca gtg atg tac 432Gly Arg Pro Gly Pro Val Gln Ile Asp Val Pro Ser Pro Val Met Tyr 130 135 140gac atg acc gac gag tcc cgg gct ggc ctg ctc gat ccg atc gcc tat 480Asp Met Thr Asp Glu Ser Arg Ala Gly Leu Leu Asp Pro Ile Ala Tyr145 150 155 160cgg gcg ccc cct cct tca gcc ggc ggc tcg caa atc aac gcc gcc gcc 528Arg Ala Pro Pro Pro Ser Ala Gly Gly Ser Gln Ile Asn Ala Ala Ala 165 170 175caa ttg ctg gcc gcc gcg act cgt ccg ctg atc atg gtc ggc tcc ggg 576Gln Leu Leu Ala Ala Ala Thr Arg Pro Leu Ile Met Val Gly Ser Gly 180 185 190gtc gac cga gct ggc gcg ggc gag gct gtg ctg cgc cta gcc gac aag 624Val Asp Arg Ala Gly Ala Gly Glu Ala Val Leu Arg Leu Ala Asp Lys 195 200 205ctg ggt tgc ggc gtc atc gcc agc ctg gcg ggc cgg tcg gcc gtc cct 672Leu Gly Cys Gly Val Ile Ala Ser Leu Ala Gly Arg Ser Ala Val Pro 210 215 220caa gat cac ccc ctc cac ctg cac gcc tat ggc gcc ggc gct gat cag 720Gln Asp His Pro Leu His Leu His Ala Tyr Gly Ala Gly Ala Asp Gln225 230 235 240gcg cga cgc gaa gcc gac gtc atc ctg gcg ctg ggc acg cgc ctg gga 768Ala Arg Arg Glu Ala Asp Val Ile Leu Ala Leu Gly Thr Arg Leu Gly 245 250 255aac atc gac acg ccc ttc gac cgc tat tgg ggc tca tcg gag ggg cac 816Asn Ile Asp Thr Pro Phe Asp Arg Tyr Trp Gly Ser Ser Glu Gly His 260 265 270aag ctg atc cag gtc gat atc gac ccc cgc aat ttg ggc gcc tca cgt 864Lys Leu Ile Gln Val Asp Ile Asp Pro Arg Asn Leu Gly Ala Ser Arg 275 280 285ccg ttg acg cta ggc atc gtt tcg gac gcg ggc agc ctc gtg gaa ggc 912Pro Leu Thr Leu Gly Ile Val Ser Asp Ala Gly Ser Leu Val Glu Gly 290 295 300ctc ctc gag gcc ctc gag aac gcg ccc acg cgc tcg ggc gcg gac gtc 960Leu Leu Glu Ala Leu Glu Asn Ala Pro Thr Arg Ser Gly Ala Asp Val305 310 315 320gac ctc acg cgc tat cgc caa atg gac gcc gaa tgg cgg cgt tcc gag 1008Asp Leu Thr Arg Tyr Arg Gln Met Asp Ala Glu Trp Arg Arg Ser Glu 325 330 335ttc gct cat atc gag gcc cat ggc ggt cca agt cct cac ccg gct gag 1056Phe Ala His Ile Glu Ala His Gly Gly Pro Ser Pro His Pro Ala Glu 340 345 350gtt atg cag acg gtg gga gag gtt ttc ggt ccc gat gcg gtg tac gtc 1104Val Met Gln Thr Val Gly Glu Val Phe Gly Pro Asp Ala Val Tyr Val 355 360 365gcc gat ggc ggt ttc acg agc ctt tgg gct cac ttt atg ttg ccc tcg 1152Ala Asp Gly Gly Phe Thr Ser Leu Trp Ala His Phe Met Leu Pro Ser 370 375 380acc aga ccg cgc tcg tac ctg aac att ctt gag atg ggg atg ctg ggc 1200Thr Arg Pro Arg Ser Tyr Leu Asn Ile Leu Glu Met Gly Met Leu Gly385 390 395 400acc ggc ata ccg tct gcg atc ggc gcg ggt ctc gga agc ccg gat cgc 1248Thr Gly Ile Pro Ser Ala Ile Gly Ala Gly Leu Gly Ser Pro Asp Arg 405 410 415cag atc gtc tgc gtt act ggc gac ggc gcg gcc ggc ttc cat tgt atg 1296Gln Ile Val Cys Val Thr Gly Asp Gly Ala Ala Gly Phe His Cys Met 420 425 430gaa ctg cag tcc gcc gtc cgc gag gac gtc aag gtg acc gtc gtc gtc 1344Glu Leu Gln Ser Ala Val Arg Glu Asp Val Lys Val Thr Val Val Val 435 440 445ctg gcc gaa ggg tcg tgg tcg atg gag gtc ccg aat gag cag gcg cgc 1392Leu Ala Glu Gly Ser Trp Ser Met Glu Val Pro Asn Glu Gln Ala Arg 450 455 460tac ggc agg acc ttc ggc acc gag atg ggc ccc gtc ctc tgg gaa agg 1440Tyr Gly Arg Thr Phe Gly Thr Glu Met Gly Pro Val Leu Trp Glu Arg465 470 475 480ttg gcc gaa agc ctg gga tgc ttc ggc ttc aag gcc gag acc gcg ccg 1488Leu Ala Glu Ser Leu Gly Cys Phe Gly Phe Lys Ala Glu Thr Ala Pro 485 490 495gat ctg cgg ccc gct ctg agc gcg gcg cgc gat gcg ctc gga ccg gcc 1536Asp Leu Arg Pro Ala Leu Ser Ala Ala Arg Asp Ala Leu Gly Pro Ala 500 505 510ctt gtg cgg gtc cgg aca gat agg gct gcg aac ctg gcc ttt ccc ccg 1584Leu Val Arg Val Arg Thr Asp Arg Ala Ala Asn Leu Ala Phe Pro Pro 515 520 525tcg atc gcc atg cgc ttc cac gag ggc tat cag ggc ctg acc ggt tga 1632Ser Ile Ala Met Arg Phe His Glu Gly Tyr Gln Gly Leu Thr Gly 530 535 540 15 543 PRT Bacterium 2412.1 15Met Ser Pro Glu Val Asp Pro Leu Leu Ala Ala Leu Asp Asp Asn Gly 1 5 10 15Ile Arg Phe Ile Pro Val Arg His Glu Ala Ala Ala Ala Tyr Met Ala 20 25 30Glu Gly Leu Tyr Lys Thr Thr Gly Gln Val Ala Ala Thr Val Thr Asn 35 40 45Pro Gly Pro Gly Thr Ala Asn Leu Leu Pro Gly Leu Val Thr Ala Lys 50 55 60His Glu Gly Val Pro Met Ile Ala Ile Thr Ala Gln His His Gly Gly 65 70 75 80Val Val Tyr Pro Ala Thr Pro Ser Thr Phe Gln Gly Ala Asp Gln Leu 85 90 95Glu Leu Leu Arg Pro Ala Val Lys Trp Gly Ala Pro Ile His Thr Trp 100 105 110Gln Arg Ile Gly Glu Val Thr Arg Met Ala Phe Arg Glu Met Trp Ala 115 120 125Gly Arg Pro Gly Pro Val Gln Ile Asp Val Pro Ser Pro Val Met Tyr 130 135 140Asp Met Thr Asp Glu Ser Arg Ala Gly Leu Leu Asp Pro Ile Ala Tyr145 150 155 160Arg Ala Pro Pro Pro Ser Ala Gly Gly Ser Gln Ile Asn Ala Ala Ala 165 170 175Gln Leu Leu Ala Ala Ala Thr Arg Pro Leu Ile Met Val Gly Ser Gly 180 185 190Val Asp Arg Ala Gly Ala Gly Glu Ala Val Leu Arg Leu Ala Asp Lys 195 200 205Leu Gly Cys Gly Val Ile Ala Ser Leu Ala Gly Arg Ser Ala Val Pro 210 215 220Gln Asp His Pro Leu His Leu His Ala Tyr Gly Ala Gly Ala Asp Gln225 230 235 240Ala Arg Arg Glu Ala Asp Val Ile Leu Ala Leu Gly Thr Arg Leu Gly 245 250 255Asn Ile Asp Thr Pro Phe Asp Arg Tyr Trp Gly Ser Ser Glu Gly His 260 265 270Lys Leu Ile Gln Val Asp Ile Asp Pro Arg Asn Leu Gly Ala Ser Arg 275 280 285Pro Leu Thr Leu Gly Ile Val Ser Asp Ala Gly Ser Leu Val Glu Gly 290 295 300Leu Leu Glu Ala Leu Glu Asn Ala Pro Thr Arg Ser Gly Ala Asp Val305 310 315 320Asp Leu Thr Arg Tyr Arg Gln Met Asp Ala Glu Trp Arg Arg Ser Glu 325 330 335Phe Ala His Ile Glu Ala His Gly Gly Pro Ser Pro His Pro Ala Glu 340 345 350Val Met Gln Thr Val Gly Glu Val Phe Gly Pro Asp Ala Val Tyr Val 355 360 365Ala Asp Gly Gly Phe Thr Ser Leu Trp Ala His Phe Met Leu Pro Ser 370 375 380Thr Arg Pro Arg Ser Tyr Leu Asn Ile Leu Glu Met Gly Met Leu Gly385 390 395 400Thr Gly Ile Pro Ser Ala Ile Gly Ala Gly Leu Gly Ser Pro Asp Arg 405 410 415Gln Ile Val Cys Val Thr Gly Asp Gly Ala Ala Gly Phe His Cys Met 420 425 430Glu Leu Gln Ser Ala Val Arg Glu Asp Val Lys Val Thr Val Val Val 435 440 445Leu Ala Glu Gly Ser Trp Ser Met Glu Val Pro Asn Glu Gln Ala Arg 450 455 460Tyr Gly Arg Thr Phe Gly Thr Glu Met Gly Pro Val Leu Trp Glu Arg465 470 475 480Leu Ala Glu Ser Leu Gly Cys Phe Gly Phe Lys Ala Glu Thr Ala Pro 485 490 495Asp Leu Arg Pro Ala Leu Ser Ala Ala Arg Asp Ala Leu Gly Pro Ala 500 505 510Leu Val Arg Val Arg Thr Asp Arg Ala Ala Asn Leu Ala Phe Pro Pro 515 520 525Ser Ile Ala Met Arg Phe His Glu Gly Tyr Gln Gly Leu Thr Gly 530 535 540 16 2865 DNA Bacterium 2412.1 fccH Vitamin B12 Receptor 16atg ccg tcc ccc cct cgc cgt cac aaa gag agg gag gga tta gcc atg 48Met Pro Ser Pro Pro Arg Arg His Lys Glu Arg Glu Gly Leu Ala Met 1 5 10 15agc caa gca gca acc aat cag aag agc act ttc cgc ctt agc ctg ctg 96Ser Gln Ala Ala Thr Asn Gln Lys Ser Thr Phe Arg Leu Ser Leu Leu 20 25 30ggc gca gcg gcg acg tcc gtg ctt cta gcc cca agc ctc ggc cgg gcg 144Gly Ala Ala Ala Thr Ser Val Leu Leu Ala Pro Ser Leu Gly Arg Ala 35 40 45cag caa gca cct gtc gaa aac cct cgg ccc agg acg cca gga agt cag 192Gln Gln Ala Pro Val Glu Asn Pro Arg Pro Arg Thr Pro Gly Ser Gln 50 55 60cga aat cgt cgt cac ggg cag ccg cct cca gag cgg att cag cgc acc 240Arg Asn Arg Arg His Gly Gln Pro Pro Pro Glu Arg Ile Gln Arg Thr 65 70 75 80gac tcc cgt gac cgc ggc gtc tgc gat cag ctc aag gcg gcg gct cca 288Asp Ser Arg Asp Arg Gly Val Cys Asp Gln Leu Lys Ala Ala Ala Pro 85 90 95acc aac atc gcc gat ggc ctc aac caa ctt ccg gtc ttc aat aat agt 336Thr Asn Ile Ala Asp Gly Leu Asn Gln Leu Pro Val Phe Asn Asn Ser 100 105 110ctg aaa acg tcg aac ccc ggc acg acg ccg ggc acc ggc aac agc ggc 384Leu Lys Thr Ser Asn Pro Gly Thr Thr Pro Gly Thr Gly Asn Ser Gly 115 120 125cag aat ctc ctg agc ctg cgc ggc ctg ggc gcg aac cga aac ctc gtt 432Gln Asn Leu Leu Ser Leu Arg Gly Leu Gly Ala Asn Arg Asn Leu Val 130 135 140ctg ctc aac ggc aac cgc ttc gtc gcc acc aac tac acc gga tcg gtg 480Leu Leu Asn Gly Asn Arg Phe Val Ala Thr Asn Tyr Thr Gly Ser Val145 150 155 160gac gtc aat gtg ttg ccc cag gct ctc gtc aag cgg gtc gac gtg gtg 528Asp Val Asn Val Leu Pro Gln Ala Leu Val Lys Arg Val Asp Val Val 165 170 175acc ggc ggc gct tcg gcc gcc tac gga tcc gat gcc gta tcc ggc gta 576Thr Gly Gly Ala Ser Ala Ala Tyr Gly Ser Asp Ala Val Ser Gly Val 180 185 190atc aac ttc gtc ctc gac gag gat ttc gag ggc cta aag gcg aac gtc 624Ile Asn Phe Val Leu Asp Glu Asp Phe Glu Gly Leu Lys Ala Asn Val 195 200 205cag acc ggc gtc tcg agc cgc aac gat ctg gcc tcg gtg ggc gga tcg 672Gln Thr Gly Val Ser Ser Arg Asn Asp Leu Ala Ser Val Gly Gly Ser 210 215 220ctg gcg gcc ggc aag tcg ttt gcc caa ggg cgc gca cat ctc ctt gcc 720Leu Ala Ala Gly Lys Ser Phe Ala Gln Gly Arg Ala His Leu Leu Ala225 230 235 240gcg gtc gaa tac tat cac gag gac ggc att cgc gcc gat cag gcc acc 768Ala Val Glu Tyr Tyr His Glu Asp Gly Ile Arg Ala Asp Gln Ala Thr 245 250 255gat cgg gcg tgg tat gat cgc gcg gcg ggg caa tat ccc gtg ccc ggc 816Asp Arg Ala Trp Tyr Asp Arg Ala Ala Gly Gln Tyr Pro Val Pro Gly 260 265 270gcc ccg acg ggc gtc acg gtg gtt ccc gac atc cga agc tcg cgc ggc 864Ala Pro Thr Gly Val Thr Val Val Pro Asp Ile Arg Ser Ser Arg Gly 275 280 285gcc tac ggc ggc ctg atc aca tcg ggt ccg ctg aag ggc gtc acc ttt 912Ala Tyr Gly Gly Leu Ile Thr Ser Gly Pro Leu Lys Gly Val Thr Phe 290 295 300ctt ccc ggc ggt acg ctt gcg acc ttt aac tac gga agc ttt acc agc 960Leu Pro Gly Gly Thr Leu Ala Thr Phe Asn Tyr Gly Ser Phe Thr Ser305 310 315 320agt tcc ttc cag agc ggc ggc gac ggt ccg cgc gtc aat ctc ggc ttc 1008Ser Ser Phe Gln Ser Gly Gly Asp Gly Pro Arg Val Asn Leu Gly Phe 325 330 335gct ccg gac cag cgg cgc tac aat ggc ttc ctc cgc ggc gag ttc gag 1056Ala Pro Asp Gln Arg Arg Tyr Asn Gly Phe Leu Arg Gly Glu Phe Glu 340 345 350gcc tca gaa cgc gtc aag ctc tat gcg gaa ggc acc tat gcc tat agc 1104Ala Ser Glu Arg Val Lys Leu Tyr Ala Glu Gly Thr Tyr Ala Tyr Ser 355 360 365cac acc aac ctc ggc gcc ttc gtc aac cag ttt gtc ggc agc gcg aac 1152His Thr Asn Leu Gly Ala Phe Val Asn Gln Phe Val Gly Ser Ala Asn 370 375 380gcc ttc acg atc ttc cgc gac aac gcg ttc ctc ccg acc gca ctc ggc 1200Ala Phe Thr Ile Phe Arg Asp Asn Ala Phe Leu Pro Thr Ala Leu Gly385 390 395 400gca ctg atg gac acc aac cgg ctg acg tcg gtt tcc gtc ggc cgg ttc 1248Ala Leu Met Asp Thr Asn Arg Leu Thr Ser Val Ser Val Gly Arg Phe 405 410 415gcc ggc gag ttc ccg ctg gtc gag atc gag tcc tac gcc aag gtg cgt 1296Ala Gly Glu Phe Pro Leu Val Glu Ile Glu Ser Tyr Ala Lys Val Arg 420 425 430cgc gga gct gcc ggc ttc cgg gcg gac ctc aac gac acc tgg aag ctc 1344Arg Gly Ala Ala Gly Phe Arg Ala Asp Leu Asn Asp Thr Trp Lys Leu 435 440 445gac ggt tcg atc tcc tac ggc cgc acg aac ctg gag ctt cgc gaa aac 1392Asp Gly Ser Ile Ser Tyr Gly Arg Thr Asn Leu Glu Leu Arg Glu Asn 450 455 460aac ctg tcg atc aac cgc aat ctc tat gca gcg gtc gac gcc gtg aag 1440Asn Leu Ser Ile Asn Arg Asn Leu Tyr Ala Ala Val Asp Ala Val Lys465 470 475 480gac ccg acg gga aag atc gtc tgc cgt tcg acc ctt tcg ggt ctc gac 1488Asp Pro Thr Gly Lys Ile Val Cys Arg Ser Thr Leu Ser Gly Leu Asp 485 490 495gcg ggc tgc gtg ccg ctc aac att ttc ggc gcc ggc gcg cca agt gcc 1536Ala Gly Cys Val Pro Leu Asn Ile Phe Gly Ala Gly Ala Pro Ser Ala 500 505 510gcc gcg atc gac tat gtc ctc gac gac ggg gtg gcg aac ctg aaa ctg 1584Ala Ala Ile Asp Tyr Val Leu Asp Asp Gly Val Ala Asn Leu Lys Leu 515 520 525gaa cag gtc gtc gcg ggc ctg aac ctc gtc ggc gac ctt ggc cag gcg 1632Glu Gln Val Val Ala Gly Leu Asn Leu Val Gly Asp Leu Gly Gln Ala 530 535 540ttt tcg ctg ggc gcc ggg ccg atc tcg atc gcc gcg ggc ggc gaa tat 1680Phe Ser Leu Gly Ala Gly Pro Ile Ser Ile Ala Ala Gly Gly Glu Tyr545 550 555 560cgt gag gag aaa gcc aac cag acc acc gac gcc atc tcg cag gcg atc 1728Arg Glu Glu Lys Ala Asn Gln Thr Thr Asp Ala Ile Ser Gln Ala Ile 565 570 575acg tcg acc gcc gga ctg cgt gga gcg ccg gcc tct cag agc aac cgg 1776Thr Ser Thr Ala Gly Leu Arg Gly Ala Pro Ala Ser Gln Ser Asn Arg 580 585 590cct ggt ggt ttc aac ctc tac aac cct ctt ccc ttc agc ggg agc tac 1824Pro Gly Gly Phe Asn Leu Tyr Asn Pro Leu Pro Phe Ser Gly Ser Tyr 595 600 605aac atc aag gag gct tat ctt gag gtc ggg gtt ccg gtg ctc aag gac 1872Asn Ile Lys Glu Ala Tyr Leu Glu Val Gly Val Pro Val Leu Lys Asp 610 615 620agc gcg ctt ggt cga gcg ctc aac ctc aac ggc gcg gtc cgg tat gcc 1920Ser Ala Leu Gly Arg Ala Leu Asn Leu Asn Gly Ala Val Arg Tyr Ala625 630 635 640gac tat agc gtc tcg ggc ggc gtc acg acc tgg aag gtc ggc ggc gac 1968Asp Tyr Ser Val Ser Gly Gly Val Thr Thr Trp Lys Val Gly Gly Asp 645 650 655tac gag ccc gtc gat gga ctg cgg ttt cgc ctg act cgc tcg cgc gac 2016Tyr Glu Pro Val Asp Gly Leu Arg Phe Arg Leu Thr Arg Ser Arg Asp 660 665 670atc cgc ggc gcc agc ctg gtg gaa ctc tac gac ccc ggc cgt cag gct 2064Ile Arg Gly Ala Ser Leu Val Glu Leu Tyr Asp Pro Gly Arg Gln Ala 675 680 685acc ctg aac tct gtc tat cag ggc cag acg ttg cag acc cgc ttc ttc 2112Thr Leu Asn Ser Val Tyr Gln Gly Gln Thr Leu Gln Thr Arg Phe Phe 690 695 700acc gcc ggc aac ccg gat ctt cgc ccc gag cgc gcc gat acc ctg acc 2160Thr Ala Gly Asn Pro Asp Leu Arg Pro Glu Arg Ala Asp Thr Leu Thr705 710 715 720ttc ggc gtc gtg ttg cgg ccg gcc ttc gcc ccg ggg ctt cag ctt tcg 2208Phe Gly Val Val Leu Arg Pro Ala Phe Ala Pro Gly Leu Gln Leu Ser 725 730 735gcg gac cgc tat atc atc gat ctc aaa gac gcc atc gac tac ctc ctg 2256Ala Asp Arg Tyr Ile Ile Asp Leu Lys Asp Ala Ile Asp Tyr Leu Leu 740 745 750ccc cag cag gag atc gac ctc tgc gcc gcg ggc aac cag tcc atg tgc 2304Pro Gln Gln Glu Ile Asp Leu Cys Ala Ala Gly Asn Gln Ser Met Cys 755 760 765gcc ttg atc acc cgg aat gcg gac aac acg ctg acc gtc atc ggc ccc 2352Ala Leu Ile Thr Arg Asn Ala Asp Asn Thr Leu Thr Val Ile Gly Pro 770 775 780aac ctg aac ctt gcg gtg cag aag gcg gcg ggt gtc gat ttg gag gcg 2400Asn Leu Asn Leu Ala Val Gln Lys Ala Ala Gly Val Asp Leu Glu Ala785 790 795 800tcc tac gtg cgc aac gtc gcg ggc gga tct ctg aac ctg cgg gcc ttg 2448Ser Tyr Val Arg Asn Val Ala Gly Gly Ser Leu Asn Leu Arg Ala Leu 805 810 815gcc aac cac cgc acg gcc gca tcc gtc acc gcc ctg ggc tcg gcg ccc 2496Ala Asn His Arg Thr Ala Ala Ser Val Thr Ala Leu Gly Ser Ala Pro 820 825 830cta caa tcg ctc ggc gaa ccc acc gcc ccc aaa tgg ctg ctc aat ctg 2544Leu Gln Ser Leu Gly Glu Pro Thr Ala Pro Lys Trp Leu Leu Asn Leu 835 840 845cag gcg cgt tac gag cgc gcc gca tgg tcg ctg ttc ctg caa gag cgc 2592Gln Ala Arg Tyr Glu Arg Ala Ala Trp Ser Leu Phe Leu Gln Glu Arg 850 855 860ttc atc tct cgc tcg gtc ttt gac gct gaa aac gtc gaa ggt gtc gac 2640Phe Ile Ser Arg Ser Val Phe Asp Ala Glu Asn Val Glu Gly Val Asp865 870 875 880acc aac ctg aac cac acc ggc gca gtc tgg tac act gac gcc acg gtg 2688Thr Asn Leu Asn His Thr Gly Ala Val Trp Tyr Thr Asp Ala Thr Val 885 890 895acc tac agc ttc gat tca ttc ggg cat aag cag cag gtt ttt gct tcg 2736Thr Tyr Ser Phe Asp Ser Phe Gly His Lys Gln Gln Val Phe Ala Ser 900 905 910gta aac aac ctc ttc gac cgt gat cca ccg gtg gcg acg gtc aat ccg 2784Val Asn Asn Leu Phe Asp Arg Asp Pro Pro Val Ala Thr Val Asn Pro 915 920 925tcc agc ttc tcg gtc ccg acg agc gca gcc tac gat ccg cgc ggc agg 2832Ser Ser Phe Ser Val Pro Thr Ser Ala Ala Tyr Asp Pro Arg Gly Arg 930 935 940tat ttc aac gtc ggg ctc cgc ttc cgc tac tga 2865Tyr Phe Asn Val Gly Leu Arg Phe Arg Tyr945 950 17 954 PRT Bacterium 2412.1 17Met Pro Ser Pro Pro Arg Arg His Lys Glu Arg Glu Gly Leu Ala Met 1 5 10 15Ser Gln Ala Ala Thr Asn Gln Lys Ser Thr Phe Arg Leu Ser Leu Leu 20 25 30Gly Ala Ala Ala Thr Ser Val Leu Leu Ala Pro Ser Leu Gly Arg Ala 35 40 45Gln Gln Ala Pro Val Glu Asn Pro Arg Pro Arg Thr Pro Gly Ser Gln 50 55 60Arg Asn Arg Arg His Gly Gln Pro Pro Pro Glu Arg Ile Gln Arg Thr 65 70 75 80Asp Ser Arg Asp Arg Gly Val Cys Asp Gln Leu Lys Ala Ala Ala Pro 85 90 95Thr Asn Ile Ala Asp Gly Leu Asn Gln Leu Pro Val Phe Asn Asn Ser 100 105 110Leu Lys Thr Ser Asn Pro Gly Thr Thr Pro Gly Thr Gly Asn Ser Gly 115 120 125Gln Asn Leu Leu Ser Leu Arg Gly Leu Gly Ala Asn Arg Asn Leu Val 130 135 140Leu Leu Asn Gly Asn Arg Phe Val Ala Thr Asn Tyr Thr Gly Ser Val145 150 155 160Asp Val Asn Val Leu Pro Gln Ala Leu Val Lys Arg Val Asp Val Val 165 170 175Thr Gly Gly Ala Ser Ala Ala Tyr Gly Ser Asp Ala Val Ser Gly Val 180 185 190Ile Asn Phe Val Leu Asp Glu Asp Phe Glu Gly Leu Lys Ala Asn Val 195 200 205Gln Thr Gly Val Ser Ser Arg Asn Asp Leu Ala Ser Val Gly Gly Ser 210 215 220Leu Ala Ala Gly Lys Ser Phe Ala Gln Gly Arg Ala His Leu Leu Ala225 230 235 240Ala Val Glu Tyr Tyr His Glu Asp Gly Ile Arg Ala Asp Gln Ala Thr 245 250 255Asp Arg Ala Trp Tyr Asp Arg Ala Ala Gly Gln Tyr Pro Val Pro Gly 260 265 270Ala Pro Thr Gly Val Thr Val Val Pro Asp Ile Arg Ser Ser Arg Gly 275 280 285Ala Tyr Gly Gly Leu Ile Thr Ser Gly Pro Leu Lys Gly Val Thr Phe 290 295 300Leu Pro Gly Gly Thr Leu Ala Thr Phe Asn Tyr Gly Ser Phe Thr Ser305 310 315 320Ser Ser Phe Gln Ser Gly Gly Asp Gly Pro Arg Val Asn Leu Gly Phe 325 330 335Ala Pro Asp Gln Arg Arg Tyr Asn Gly Phe Leu Arg Gly Glu Phe Glu 340 345 350Ala Ser Glu Arg Val Lys Leu Tyr Ala Glu Gly Thr Tyr Ala Tyr Ser 355 360 365His Thr Asn Leu Gly Ala Phe Val Asn Gln Phe Val Gly Ser Ala Asn 370 375 380Ala Phe Thr Ile Phe Arg Asp Asn Ala Phe Leu Pro Thr Ala Leu Gly385 390 395 400Ala Leu Met Asp Thr Asn Arg Leu Thr Ser Val Ser Val Gly Arg Phe 405 410 415Ala Gly Glu Phe Pro Leu Val Glu Ile Glu Ser Tyr Ala Lys Val Arg 420 425 430Arg Gly Ala Ala Gly Phe Arg Ala Asp Leu Asn Asp Thr Trp Lys Leu 435 440 445Asp Gly Ser Ile Ser Tyr Gly Arg Thr Asn Leu Glu Leu Arg Glu Asn 450 455 460Asn Leu Ser Ile Asn Arg Asn Leu Tyr Ala Ala Val Asp Ala Val Lys465 470 475 480Asp Pro Thr Gly Lys Ile Val Cys Arg Ser Thr Leu Ser Gly Leu Asp 485 490 495Ala Gly Cys Val Pro Leu Asn Ile Phe Gly Ala Gly Ala Pro Ser Ala 500 505 510Ala Ala Ile Asp Tyr Val Leu Asp Asp Gly Val Ala Asn Leu Lys Leu 515 520 525Glu Gln Val Val Ala Gly Leu Asn Leu Val Gly Asp Leu Gly Gln Ala 530 535 540Phe Ser Leu Gly Ala Gly Pro Ile Ser Ile Ala Ala Gly Gly Glu Tyr545 550 555 560Arg Glu Glu Lys Ala Asn Gln Thr Thr Asp Ala Ile Ser Gln Ala Ile 565 570 575Thr Ser Thr Ala Gly Leu Arg Gly Ala Pro Ala Ser Gln Ser Asn Arg 580 585 590Pro Gly Gly Phe Asn Leu Tyr Asn Pro Leu Pro Phe Ser Gly Ser Tyr 595 600 605Asn Ile Lys Glu Ala Tyr Leu Glu Val Gly Val Pro Val Leu Lys Asp 610 615 620Ser Ala Leu Gly Arg Ala Leu Asn Leu Asn Gly Ala Val Arg Tyr Ala625 630 635 640Asp Tyr Ser Val Ser Gly Gly Val Thr Thr Trp Lys Val Gly Gly Asp 645 650 655Tyr Glu Pro Val Asp Gly Leu Arg Phe Arg Leu Thr Arg Ser Arg Asp 660 665 670Ile Arg Gly Ala Ser Leu Val Glu Leu Tyr Asp Pro Gly Arg Gln Ala 675 680 685Thr Leu Asn Ser Val Tyr Gln Gly Gln Thr Leu Gln Thr Arg Phe Phe 690 695 700Thr Ala Gly Asn Pro Asp Leu Arg Pro Glu Arg Ala Asp Thr Leu Thr705 710 715 720Phe Gly Val Val Leu Arg Pro Ala Phe Ala Pro Gly Leu Gln Leu Ser 725 730 735Ala Asp Arg Tyr Ile Ile Asp Leu Lys Asp Ala Ile Asp Tyr Leu Leu 740 745 750Pro Gln Gln Glu Ile Asp Leu Cys Ala Ala Gly Asn Gln Ser Met Cys 755 760 765Ala Leu Ile Thr Arg Asn Ala Asp Asn Thr Leu Thr Val Ile Gly Pro 770 775 780Asn Leu Asn Leu Ala Val Gln Lys Ala Ala Gly Val Asp Leu Glu Ala785 790 795 800Ser Tyr Val Arg Asn Val Ala Gly Gly Ser Leu Asn Leu Arg Ala Leu 805 810 815Ala Asn His Arg Thr Ala Ala Ser Val Thr Ala Leu Gly Ser Ala Pro 820 825 830Leu Gln Ser Leu Gly Glu Pro Thr Ala Pro Lys Trp Leu Leu Asn Leu 835 840 845Gln Ala Arg Tyr Glu Arg Ala Ala Trp Ser Leu Phe Leu Gln Glu Arg 850 855 860Phe Ile Ser Arg Ser Val Phe Asp Ala Glu Asn Val Glu Gly Val Asp865 870 875 880Thr Asn Leu Asn His Thr Gly Ala Val Trp Tyr Thr Asp Ala Thr Val 885 890 895Thr Tyr Ser Phe Asp Ser Phe Gly His Lys Gln Gln Val Phe Ala Ser 900 905 910Val Asn Asn Leu Phe Asp Arg Asp Pro Pro Val Ala Thr Val Asn Pro 915 920 925Ser Ser Phe Ser Val Pro Thr Ser Ala Ala Tyr Asp Pro Arg Gly Arg 930 935 940Tyr Phe Asn Val Gly Leu Arg Phe Arg Tyr945 950 18 1302 DNA Bacterium 2412.1 fccI Permease 18atg cgt gtt ttc gta aat cgc gga cga tgt cag aat tgc tct caa act 48Met Arg Val Phe Val Asn Arg Gly Arg Cys Gln Asn Cys Ser Gln Thr 1 5 10 15att atc gat att tgc tgg gtg tgt gga gcg cag ccg ctc ccc cac aga 96Ile Ile Asp Ile Cys Trp Val Cys Gly Ala Gln Pro Leu Pro His Arg 20 25 30tgc gaa ttc cct tca cct tct cga cga cag ttg gag cct atg tcc ctg 144Cys Glu Phe Pro Ser Pro Ser Arg Arg Gln Leu Glu Pro Met Ser Leu 35 40 45acc ctg ccc cgg aat acc gga cct cgc cat cac cag gcc gtg tcc gtc 192Thr Leu Pro Arg Asn Thr Gly Pro Arg His His Gln Ala Val Ser Val 50 55 60gta tcg gcc act atg ctg atc ggc acg gcc tcg atg ctg gtg atg ggc 240Val Ser Ala Thr Met Leu Ile Gly Thr Ala Ser Met Leu Val Met Gly 65 70 75 80gtc gaa ccc atc ctg ctc ggg ggt ctc gcc aac gcc ggg cgg atc agc 288Val Glu Pro Ile Leu Leu Gly Gly Leu Ala Asn Ala Gly Arg Ile Ser 85 90 95gag gcg ggc gta ggt cag gcc gcc atg atc gaa gtc ttc gcc ctg gcc 336Glu Ala Gly Val Gly Gln Ala Ala Met Ile Glu Val Phe Ala Leu Ala 100 105 110gca ggc tcg acg gcg ggt ccg ttc ctg atg aac ctc ggc cac atg cgc 384Ala Gly Ser Thr Ala Gly Pro Phe Leu Met Asn Leu Gly His Met Arg 115 120 125gcc aag gtg gcc gct gcc tcc ctg ctc ctt gcg atc atc aac ctc gcc 432Ala Lys Val Ala Ala Ala Ser Leu Leu Leu Ala Ile Ile Asn Leu Ala 130 135 140atc tac tgg gcc gca tcc ccg gcc acg atc ctg gtc caa cgc ggc gcg 480Ile Tyr Trp Ala Ala Ser Pro Ala Thr Ile Leu Val Gln Arg Gly Ala145 150 155 160gcc ggg ttg ctg gaa ggg cta ttg ctg ggc gcg gcc ggc gcc atc ctt 528Ala Gly Leu Leu Glu Gly Leu Leu Leu Gly Ala Ala Gly Ala Ile Leu 165 170 175acc cac aac gac cgg cct gag cgc atg agc ggg ctg ctg ctc ggt ctt 576Thr His Asn Asp Arg Pro Glu Arg Met Ser Gly Leu Leu Leu Gly Leu 180 185 190tcc acg atc ccc cag gtg atc gcg gcc tat ttg ctg ccg atc tgg gtg 624Ser Thr Ile Pro Gln Val Ile Ala Ala Tyr Leu Leu Pro Ile Trp Val 195 200 205atc ccg cgg ttt ggc gtg gat gcc ggg ttc gcg gtc ttg gcg ggc gtt 672Ile Pro Arg Phe Gly Val Asp Ala Gly Phe Ala Val Leu Ala Gly Val 210 215 220gcg atg gcc tcc tgc ctc gtt gcg ccg gcg atc gtc gac cac gtg cct 720Ala Met Ala Ser Cys Leu Val Ala Pro Ala Ile Val Asp His Val Pro225 230 235 240gct cct acg gga tcg cac cat ggt cgc gta gtc gtc tct ccg gcc ctg 768Ala Pro Thr Gly Ser His His Gly Arg Val Val Val Ser Pro Ala Leu 245 250 255atg gtc gtg gcg ctc gcg gcg ttt ctt caa aac gcc ggc atc ggg gcg 816Met Val Val Ala Leu Ala Ala Phe Leu Gln Asn Ala Gly Ile Gly Ala 260 265 270gca tgg aac tac ctg gag cgc ctg gcc gcg caa cac cat ttc gcc ccg 864Ala Trp Asn Tyr Leu Glu Arg Leu Ala Ala Gln His His Phe Ala Pro 275 280 285gcc acg gtc ggc gcc gcg atc gcg ggc agc ctg gcc ttc cag gtg gcg 912Ala Thr Val Gly Ala Ala Ile Ala Gly Ser Leu Ala Phe Gln Val Ala 290 295 300ggt gct ctt gca gca tcc tgg ctc ggt gcg cgc gtg cac gcc cgt acg 960Gly Ala Leu Ala Ala Ser Trp Leu Gly Ala Arg Val His Ala Arg Thr305 310 315 320gtt ctg gcc gcc ggc gca gtg ctg cag gcc ggt ctt gtc atc ggc ctg 1008Val Leu Ala Ala Gly Ala Val Leu Gln Ala Gly Leu Val Ile Gly Leu 325 330 335ctt cac gcc ggc acg ccc gtc gcg ctc atc gca agc gct tgt ggc ttt 1056Leu His Ala Gly Thr Pro Val Ala Leu Ile Ala Ser Ala Cys Gly Phe 340 345 350ggc ctg ttc tgg ctg gcg ctg cag ccg ttt ctg gtg gcg gac gtg atc 1104Gly Leu Phe Trp Leu Ala Leu Gln Pro Phe Leu Val Ala Asp Val Ile 355 360 365gcc ttg gag ccc agc agg acc gcc gcc gta ctg ctc gcc cct ctc gcg 1152Ala Leu Glu Pro Ser Arg Thr Ala Ala Val Leu Leu Ala Pro Leu Ala 370 375 380ttg gtg ggc ttc agc gcc ggt ccg ttg gcg gcc tcg ttc gtc att gac 1200Leu Val Gly Phe Ser Ala Gly Pro Leu Ala Ala Ser Phe Val Ile Asp385 390 395 400gat aat cgg gtg gga ggc gca ttc cag gtt tcg gcc ggc tcc tcg ttg 1248Asp Asn Arg Val Gly Gly Ala Phe Gln Val Ser Ala Gly Ser Ser Leu 405 410 415cgg cgg cgg cgc tct acg tcg tgg ccg gcc cct ggc ggc ggg cgg cta 1296Arg Arg Arg Arg Ser Thr Ser Trp Pro Ala Pro Gly Gly Gly Arg Leu 420 425 430ctt taa 1302Leu 19 433 PRT Bacterium 2412.1 19Met Arg Val Phe Val Asn Arg Gly Arg Cys Gln Asn Cys Ser Gln Thr 1 5 10 15Ile Ile Asp Ile Cys Trp Val Cys Gly Ala Gln Pro Leu Pro His Arg 20 25 30Cys Glu Phe Pro Ser Pro Ser Arg Arg Gln Leu Glu Pro Met Ser Leu 35 40 45Thr Leu Pro Arg Asn Thr Gly Pro Arg His His Gln Ala Val Ser Val 50 55 60Val Ser Ala Thr Met Leu Ile Gly Thr Ala Ser Met Leu Val Met Gly 65 70 75 80Val Glu Pro Ile Leu Leu Gly Gly Leu Ala Asn Ala Gly Arg Ile Ser 85 90 95Glu Ala Gly Val Gly Gln Ala Ala Met Ile Glu Val Phe Ala Leu Ala 100 105 110Ala Gly Ser Thr Ala Gly Pro Phe Leu Met Asn Leu Gly His Met Arg 115 120 125Ala Lys Val Ala Ala Ala Ser Leu Leu Leu Ala Ile Ile Asn Leu Ala 130 135 140Ile Tyr Trp Ala Ala Ser Pro Ala Thr Ile Leu Val Gln Arg Gly Ala145 150 155 160Ala Gly Leu Leu Glu Gly Leu Leu Leu Gly Ala Ala Gly Ala Ile Leu 165 170 175Thr His Asn Asp Arg Pro Glu Arg Met Ser Gly Leu Leu Leu Gly Leu 180 185 190Ser Thr Ile Pro Gln Val Ile Ala Ala Tyr Leu Leu Pro Ile Trp Val 195 200 205Ile Pro Arg Phe Gly Val Asp Ala Gly Phe Ala Val Leu Ala Gly Val 210 215 220Ala Met Ala Ser Cys Leu Val Ala Pro Ala Ile Val Asp His Val Pro225 230 235 240Ala Pro Thr Gly Ser His His Gly Arg Val Val Val Ser Pro Ala Leu 245 250 255Met Val Val Ala Leu Ala Ala Phe Leu Gln Asn Ala Gly Ile Gly Ala 260 265 270Ala Trp Asn Tyr Leu Glu Arg Leu Ala Ala Gln His His Phe Ala Pro 275 280 285Ala Thr Val Gly Ala Ala Ile Ala Gly Ser Leu Ala Phe Gln Val Ala 290 295 300Gly Ala Leu Ala Ala Ser Trp Leu Gly Ala Arg Val His Ala Arg Thr305 310 315 320Val Leu Ala Ala Gly Ala Val Leu Gln Ala Gly Leu Val Ile Gly Leu 325 330 335Leu His Ala Gly Thr Pro Val Ala Leu Ile Ala Ser Ala Cys Gly Phe 340 345 350Gly Leu Phe Trp Leu Ala Leu Gln Pro Phe Leu Val Ala Asp Val Ile 355 360 365Ala Leu Glu Pro Ser Arg Thr Ala Ala Val Leu Leu Ala Pro Leu Ala 370 375 380Leu Val Gly Phe Ser Ala Gly Pro Leu Ala Ala Ser Phe Val Ile Asp385 390 395 400Asp Asn Arg Val Gly Gly Ala Phe Gln Val Ser Ala Gly Ser Ser Leu 405 410 415Arg Arg Arg Arg Ser Thr Ser Trp Pro Ala Pro Gly Gly Gly Arg Leu 420 425 430Leu 20 912 DNA Bacterium 2412.1 fccJ Regulatory Protein 20atg ttg aac tgc gat cta ttg gat ctg cgc gcc ttc gtc gcg gtc cac 48Met Leu Asn Cys Asp Leu Leu Asp Leu Arg Ala Phe Val Ala Val His 1 5 10 15gaa acc cgc agc ttc atc cgc gcg gcg cat ctg ctc ggc ctt tcc cag 96Glu Thr Arg Ser Phe Ile Arg Ala Ala His Leu Leu Gly Leu Ser Gln 20 25 30ccc gcg ctc agt cgc cgg atc caa cgc ttg gag gga ctg gtc ggc ggc 144Pro Ala Leu Ser Arg Arg Ile Gln Arg Leu Glu Gly Leu Val Gly Gly 35 40 45gct ctc ttc gac cga acc agc cgg acc atg acc gag acc gcg ctt ggc 192Ala Leu Phe Asp Arg Thr Ser Arg Thr Met Thr Glu Thr Ala Leu Gly 50 55 60aag gag ctg ctg ccg gtg gcc cgc cga acg ctt gag ttt ctg gac aat 240Lys Glu Leu Leu Pro Val Ala Arg Arg Thr Leu Glu Phe Leu Asp Asn 65 70 75 80tcg ctg ttc gcc tcg ccc aag ctg cgc gaa ccg cgc tgg acc gac atc 288Ser Leu Phe Ala Ser Pro Lys Leu Arg Glu Pro Arg Trp Thr Asp Ile 85 90 95agc att ttt tgc gtg cag acc gcc gcg ttc cgc gtt ctg ccg cgc gcg 336Ser Ile Phe Cys Val Gln Thr Ala Ala Phe Arg Val Leu Pro Arg Ala 100 105 110gcc cgg cgc ttc atg gat gaa aat ccc cga ctg cgc ctg agg atc atc 384Ala Arg Arg Phe Met Asp Glu Asn Pro Arg Leu Arg Leu Arg Ile Ile 115 120 125gat gtt ccg gct gtc gaa ggc gcg gaa ctg gtg gcg cga ggg gaa gcg 432Asp Val Pro Ala Val Glu Gly Ala Glu Leu Val Ala Arg Gly Glu Ala 130 135 140gag ttc ggt atc agc atc gag agc ctg ctt ccg tcc ggc ctg cgt ttc 480Glu Phe Gly Ile Ser Ile Glu Ser Leu Leu Pro Ser Gly Leu Arg Phe145 150 155 160gag gct ctt cac gag gac ccg ttt ggc ttg gcg tgc cat cgg agc cat 528Glu Ala Leu His Glu Asp Pro Phe Gly Leu Ala Cys His Arg Ser His 165 170 175cgc ctg gcg caa agc gac gtc atc gaa tgg tcc gcg ctc cgc ggc gaa 576Arg Leu Ala Gln Ser Asp Val Ile Glu Trp Ser Ala Leu Arg Gly Glu 180 185 190aat ctt gtc gcc gtc cac cgg gcc agt cgc aac cgg acc ctg ctc gac 624Asn Leu Val Ala Val His Arg Ala Ser Arg Asn Arg Thr Leu Leu Asp 195 200 205gcc gag ctc aag cag cat gcg atc tcc ctg gac tgg cgt tac gag gtc 672Ala Glu Leu Lys Gln His Ala Ile Ser Leu Asp Trp Arg Tyr Glu Val 210 215 220ggt cac ttg acg acc gcg ctg ggg ctg atc gag tcc gag gtc ggc gtg 720Gly His Leu Thr Thr Ala Leu Gly Leu Ile Glu Ser Glu Val Gly Val225 230 235 240gcc gtc atg ccg cgg atg gtg atg ccc caa tca ggc cgc tca gaa ctg 768Ala Val Met Pro Arg Met Val Met Pro Gln Ser Gly Arg Ser Glu Leu 245 250 255gtc tgg gtt ccc ttg gtc gcc ccg gtc gtg agg cgc acg atc ggc atc 816Val Trp Val Pro Leu Val Ala Pro Val Val Arg Arg Thr Ile Gly Ile 260 265 270gtg cag cgc cgg gtg ggc gcg atg cat ccc gcc gcc gcc caa ctg ctc 864Val Gln Arg Arg Val Gly Ala Met His Pro Ala Ala Ala Gln Leu Leu 275 280 285gag cgg ttg cgg gag gaa tgg ccg acc ggc gcg ccc gcg gac gag tag 912Glu Arg Leu Arg Glu Glu Trp Pro Thr Gly Ala Pro Ala Asp Glu 290 295 300 21 303 PRT Bacterium 2412.1 21Met Leu Asn Cys Asp Leu Leu Asp Leu Arg Ala Phe Val Ala Val His 1 5 10 15Glu Thr Arg Ser Phe Ile Arg Ala Ala His Leu Leu Gly Leu Ser Gln 20 25 30Pro Ala Leu Ser Arg Arg Ile Gln Arg Leu Glu Gly Leu Val Gly Gly 35 40 45Ala Leu Phe Asp Arg Thr Ser Arg Thr Met Thr Glu Thr Ala Leu Gly 50 55 60Lys Glu Leu Leu Pro Val Ala Arg Arg Thr Leu Glu Phe Leu Asp Asn 65 70 75 80Ser Leu Phe Ala Ser Pro Lys Leu Arg Glu Pro Arg Trp Thr Asp Ile 85 90 95Ser Ile Phe Cys Val Gln Thr Ala Ala Phe Arg Val Leu Pro Arg Ala 100 105 110Ala Arg Arg Phe Met Asp Glu Asn Pro Arg Leu Arg Leu Arg Ile Ile 115 120 125Asp Val Pro Ala Val Glu Gly Ala Glu Leu Val Ala Arg Gly Glu Ala 130 135 140Glu Phe Gly Ile Ser Ile Glu Ser Leu Leu Pro Ser Gly Leu Arg Phe145 150 155 160Glu Ala Leu His Glu Asp Pro Phe Gly Leu Ala Cys His Arg Ser His 165 170 175Arg Leu Ala Gln Ser Asp Val Ile Glu Trp Ser Ala Leu Arg Gly Glu 180 185 190Asn Leu Val Ala Val His Arg Ala Ser Arg Asn Arg Thr Leu Leu Asp 195 200 205Ala Glu Leu Lys Gln His Ala Ile Ser Leu Asp Trp Arg Tyr Glu Val 210 215 220Gly His Leu Thr Thr Ala Leu Gly Leu Ile Glu Ser Glu Val Gly Val225 230 235 240Ala Val Met Pro Arg Met Val Met Pro Gln Ser Gly Arg Ser Glu Leu 245 250 255Val Trp Val Pro Leu Val Ala Pro Val Val Arg Arg Thr Ile Gly Ile 260 265 270Val Gln Arg Arg Val Gly Ala Met His Pro Ala Ala Ala Gln Leu Leu 275 280 285Glu Arg Leu Arg Glu Glu Trp Pro Thr Gly Ala Pro Ala Asp Glu 290 295 300 22 1389 DNA Bacterium 2412.1 fccK Fumarate Reductase/Aspartate oxidase 22atg ggt gcg aag ttg aag tat gac gtg gtg gtg gtc ggg ggc ggc aat 48Met Gly Ala Lys Leu Lys Tyr Asp Val Val Val Val Gly Gly Gly Asn 1 5 10 15gcg gcg atg acg gcg gcc gtc acc gcg cgg gaa gcc ggc gcg acg gtg 96Ala Ala Met Thr Ala Ala Val Thr Ala Arg Glu Ala Gly Ala Thr Val 20 25 30ctg gtg ctt gag cat gcg ccc cgg tcg atg cgc ggc ggc aac agc cgc 144Leu Val Leu Glu His Ala Pro Arg Ser Met Arg Gly Gly Asn Ser Arg 35 40 45cat acg cgc aac atg cgc acg atg cac gag gcg cca ctt gcg gtc ttg 192His Thr Arg Asn Met Arg Thr Met His Glu Ala Pro Leu Ala Val Leu 50 55 60acc ggg caa tat tcc gaa gac gaa tac tgg aac gac ctg aag cgg gtc 240Thr Gly Gln Tyr Ser Glu Asp Glu Tyr Trp Asn Asp Leu Lys Arg Val 65 70 75 80acg ggc ggg gaa acc gac gag gcc ctg gcc cgt ctg gtg atc cgc agc 288Thr Gly Gly Glu Thr Asp Glu Ala Leu Ala Arg Leu Val Ile Arg Ser 85 90 95acg acg gac gcc atc ccc ttc atg ctc cgg tgc ggc gtg cgc ttc cag 336Thr Thr Asp Ala Ile Pro Phe Met Leu Arg Cys Gly Val Arg Phe Gln 100 105 110cca tcg ctg tcg ggc acc ttg agc ctg tcg cgg acc aac gcg ttc ttc 384Pro Ser Leu Ser Gly Thr Leu Ser Leu Ser Arg Thr Asn Ala Phe Phe 115 120 125ctg ggg ggc ggc aag gct ctg gtg aac gcc tac tac gcg acc gcc gag 432Leu Gly Gly Gly Lys Ala Leu Val Asn Ala Tyr Tyr Ala Thr Ala Glu 130 135 140cgc ctg ggc gtc gac atc ctc tat gac agc gaa gtc acc gag atc gtg 480Arg Leu Gly Val Asp Ile Leu Tyr Asp Ser Glu Val Thr Glu Ile Val145 150 155 160ctc gaa ggc ggc cgg gtc cgg cgt ctg gtg gtc cgc agc cag ggg ttc 528Leu Glu Gly Gly Arg Val Arg Arg Leu Val Val Arg Ser Gln Gly Phe 165 170 175ccc atc gag gtg gag gcg cgc gcg gtg atc gcc tcg tcg ggc ggc ttc 576Pro Ile Glu Val Glu Ala Arg Ala Val Ile Ala Ser Ser Gly Gly Phe 180 185 190cag gcc aac ctg caa tgg ctg gcg aac gcc tgg ggc ccg gcg gcg tcg 624Gln Ala Asn Leu Gln Trp Leu Ala Asn Ala Trp Gly Pro Ala Ala Ser 195 200 205aat ttc atc gta cgc ggg acg ccc tac gcg acg ggc acg gtg ctg cgc 672Asn Phe Ile Val Arg Gly Thr Pro Tyr Ala Thr Gly Thr Val Leu Arg 210 215 220aac ctg ctc gac cag ggc gtg gcc tcg gtg ggc gat ccg acc cag tgc 720Asn Leu Leu Asp Gln Gly Val Ala Ser Val Gly Asp Pro Thr Gln Cys225 230 235 240cat gct gtc gcc atc gac ggg cgc gcg ccc aag tac gac ggg ggg atc 768His Ala Val Ala Ile Asp Gly Arg Ala Pro Lys Tyr Asp Gly Gly Ile 245 250 255gtc acc cga ctg gac tgc gtg ccg ttc tcg atc gtg gtc aat cgc gac 816Val Thr Arg Leu Asp Cys Val Pro Phe Ser Ile Val Val Asn Arg Asp 260 265 270ggc caa cgc ttc tac gac gag ggc gag gac atc tgg ccc aag cga tat 864Gly Gln Arg Phe Tyr Asp Glu Gly Glu Asp Ile Trp Pro Lys Arg Tyr 275 280 285gcg atc tgg ggg cgt ctg acc gcg caa cag ccc gat cag atc gcc tac 912Ala Ile Trp Gly Arg Leu Thr Ala Gln Gln Pro Asp Gln Ile Ala Tyr 290 295 300agc atc atc gac agc cga tcc gaa cga ctt ttc atg ccg tcg gtg ttt 960Ser Ile Ile Asp Ser Arg Ser Glu Arg Leu Phe Met Pro Ser Val Phe305 310 315 320ccc ccg atc aaa gcc gac tcg att tcc gaa ctc gcg gcc aag ctc ggg 1008Pro Pro Ile Lys Ala Asp Ser Ile Ser Glu Leu Ala Ala Lys Leu Gly 325 330 335ctg gag ccg gcg acg ctc gcg cag acc atc gag acg ttc aat cgc gcc 1056Leu Glu Pro Ala Thr Leu Ala Gln Thr Ile Glu Thr Phe Asn Arg Ala 340 345 350tgc caa ccc ggt cgc ttc gat ccg cag gat ctt gac ggg gtc cgc acc 1104Cys Gln Pro Gly Arg Phe Asp Pro Gln Asp Leu Asp Gly Val Arg Thr 355 360 365gag ggg atc acg ccg tgc aag tcc aat tgg gcc cgg ccg atc acc gag 1152Glu Gly Ile Thr Pro Cys Lys Ser Asn Trp Ala Arg Pro Ile Thr Glu 370 375 380ccg ccg ttc agc gca tat ccc ctg cgg ccc ggc atc acc ttc acc tac 1200Pro Pro Phe Ser Ala Tyr Pro Leu Arg Pro Gly Ile Thr Phe Thr Tyr385 390 395 400ctc ggc gtc aag gtc gat gaa cgc gcc agg gtg atc ctg gcc tcc ggc 1248Leu Gly Val Lys Val Asp Glu Arg Ala Arg Val Ile Leu Ala Ser Gly 405 410 415cag ccg aca gag aac ctg ttc gcg tct ggc gag atc atg gcc ggg agc 1296Gln Pro Thr Glu Asn Leu Phe Ala Ser Gly Glu Ile Met Ala Gly Ser 420 425 430att ctt ggg cgc ggt tac ctg gcg ggc ttc ggc atg gcg atc ggg acc 1344Ile Leu Gly Arg Gly Tyr Leu Ala Gly Phe Gly Met Ala Ile Gly Thr 435 440 445gtc ttc gga cgc att gcg ggc cgg gag gcc gca tat cat gca gca 1389Val Phe Gly Arg Ile Ala Gly Arg Glu Ala Ala Tyr His Ala Ala 450 455 460 23 463 PRT Bacterium 2412.1 23Met Gly Ala Lys Leu Lys Tyr Asp Val Val Val Val Gly Gly Gly Asn 1 5 10 15Ala Ala Met Thr Ala Ala Val Thr Ala Arg Glu Ala Gly Ala Thr Val 20 25 30Leu Val Leu Glu His Ala Pro Arg Ser Met Arg Gly Gly Asn Ser Arg 35 40 45His Thr Arg Asn Met Arg Thr Met His Glu Ala Pro Leu Ala Val Leu 50 55 60Thr Gly Gln Tyr Ser Glu Asp Glu Tyr Trp Asn Asp Leu Lys Arg Val 65 70 75 80Thr Gly Gly Glu Thr Asp Glu Ala Leu Ala Arg Leu Val Ile Arg Ser 85 90 95Thr Thr Asp Ala Ile Pro Phe Met Leu Arg Cys Gly Val Arg Phe Gln 100 105 110Pro Ser Leu Ser Gly Thr Leu Ser Leu Ser Arg Thr Asn Ala Phe Phe 115 120 125Leu Gly Gly Gly Lys Ala Leu Val Asn Ala Tyr Tyr Ala Thr Ala Glu 130 135 140Arg Leu Gly Val Asp Ile Leu Tyr Asp Ser Glu Val Thr Glu Ile Val145 150 155 160Leu Glu Gly Gly Arg Val Arg Arg Leu Val Val Arg Ser Gln Gly Phe 165 170 175Pro Ile Glu Val Glu Ala Arg Ala Val Ile Ala Ser Ser Gly Gly Phe 180 185 190Gln Ala Asn Leu Gln Trp Leu Ala Asn Ala Trp Gly Pro Ala Ala Ser 195 200 205Asn Phe Ile Val Arg Gly Thr Pro Tyr Ala Thr Gly Thr Val Leu Arg 210 215 220Asn Leu Leu Asp Gln Gly Val Ala Ser Val Gly Asp Pro Thr Gln Cys225 230 235 240His Ala Val Ala Ile Asp Gly Arg Ala Pro Lys Tyr Asp Gly Gly Ile 245 250 255Val Thr Arg Leu Asp Cys Val Pro Phe Ser Ile Val Val Asn Arg Asp 260 265 270Gly Gln Arg Phe Tyr Asp Glu Gly Glu Asp Ile Trp Pro Lys Arg Tyr 275 280 285Ala Ile Trp Gly Arg Leu Thr Ala Gln Gln Pro Asp Gln Ile Ala Tyr 290 295 300Ser Ile Ile Asp Ser Arg Ser Glu Arg Leu Phe Met Pro Ser Val Phe305 310 315 320Pro Pro Ile Lys Ala Asp Ser Ile Ser Glu Leu Ala Ala Lys Leu Gly 325 330 335Leu Glu Pro Ala Thr Leu Ala Gln Thr Ile Glu Thr Phe Asn Arg Ala 340 345 350Cys Gln Pro Gly Arg Phe Asp Pro Gln Asp Leu Asp Gly Val Arg Thr 355 360 365Glu Gly Ile Thr Pro Cys Lys Ser Asn Trp Ala Arg Pro Ile Thr Glu 370 375 380Pro Pro Phe Ser Ala Tyr Pro Leu Arg Pro Gly Ile Thr Phe Thr Tyr385 390 395 400Leu Gly Val Lys Val Asp Glu Arg Ala Arg Val Ile Leu Ala Ser Gly 405 410 415Gln Pro Thr Glu Asn Leu Phe Ala Ser Gly Glu Ile Met Ala Gly Ser 420 425 430Ile Leu Gly Arg Gly Tyr Leu Ala Gly Phe Gly Met Ala Ile Gly Thr 435 440 445Val Phe Gly Arg Ile Ala Gly Arg Glu Ala Ala Tyr His Ala Ala 450 455 460 24 2403 DNA Bacterium 2412.1 fccL TonB-dependent Receptor 24atg cct agg gat acg aca ccg aat ttc gct ccg gtc acc acg gca aaa 48Met Pro Arg Asp Thr Thr Pro Asn Phe Ala Pro Val Thr Thr Ala Lys 1 5 10 15gag ggc cgc cga cac cga ggc agc acc gcc tta cga agg ctc atg ctg 96Glu Gly Arg Arg His Arg Gly Ser Thr Ala Leu Arg Arg Leu Met Leu 20 25 30acg gcg gcc ggc agc gcc ctg gtg ctg ggt ctt gcg ccc aag gcg ctc 144Thr Ala Ala Gly Ser Ala Leu Val Leu Gly Leu Ala Pro Lys Ala Leu 35 40 45gcg cag gtg gcg gtt ccg ccg gct ggt cac gag gcg tcg cag gag gtg 192Ala Gln Val Ala Val Pro Pro Ala Gly His Glu Ala Ser Gln Glu Val 50 55 60cag gag atc gtc gtc acc gcg cag cgc cgc agc gag aac att cag aat 240Gln Glu Ile Val Val Thr Ala Gln Arg Arg Ser Glu Asn Ile Gln Asn 65 70 75 80gtg ccg gtc tcg gtg cag gcg ctg tcg gca gcg cag ctc gag cgc gaa 288Val Pro Val Ser Val Gln Ala Leu Ser Ala Ala Gln Leu Glu Arg Glu 85 90 95ggg atc aaa cag acc agc gat atc gcc cga gtg acg ccc aac gtc acc 336Gly Ile Lys Gln Thr Ser Asp Ile Ala Arg Val Thr Pro Asn Val Thr 100 105 110atc gcc atg ccc aac ggc gaa ggc aac cag ccg gcg gtg acg atc cgc 384Ile Ala Met Pro Asn Gly Glu Gly Asn Gln Pro Ala Val Thr Ile Arg 115 120 125ggc atc ggc ctc aac gac ttc aat tcc aac aac gcc ggc ccg aac gcg 432Gly Ile Gly Leu Asn Asp Phe Asn Ser Asn Asn Ala Gly Pro Asn Ala 130 135 140atc tat gtc gac gat gtc tat atc agc gcc ccg tcg gcc cag acc ttc 480Ile Tyr Val Asp Asp Val Tyr Ile Ser Ala Pro Ser Ala Gln Thr Phe145 150 155 160gga atc ttc gac atc aac cag atc cag gtt ctc aaa gga ccg caa ggt 528Gly Ile Phe Asp Ile Asn Gln Ile Gln Val Leu Lys Gly Pro Gln Gly 165 170 175acg ctc tat ggg cgc aac tcc agc ggt ggg gcc ttg gtg ttc acg tcc 576Thr Leu Tyr Gly Arg Asn Ser Ser Gly Gly Ala Leu Val Phe Thr Ser 180 185 190aga gcg ccg agc caa gac ttc gcc gcg gac gcc cat ttc gat tac ggc 624Arg Ala Pro Ser Gln Asp Phe Ala Ala Asp Ala His Phe Asp Tyr Gly 195 200 205agc tac aac acc tat caa ctg caa gcc ggc gtc ggc ggc cct ctg agc 672Ser Tyr Asn Thr Tyr Gln Leu Gln Ala Gly Val Gly Gly Pro Leu Ser 210 215 220gat cag cta agc gcc cgc ctg gcc ttc gtc gtc aac cac tcc gac ggg 720Asp Gln Leu Ser Ala Arg Leu Ala Phe Val Val Asn His Ser Asp Gly225 230 235 240ttc atg cac aac acg ctg acg ggc ggt tcg gcg tcg ggc acg gac aat 768Phe Met His Asn Thr Leu Thr Gly Gly Ser Ala Ser Gly Thr Asp Asn 245 250 255cag gcc gtc agg ctg caa ctg ctc tac cga cct aat gac agg ctg aaa 816Gln Ala Val Arg Leu Gln Leu Leu Tyr Arg Pro Asn Asp Arg Leu Lys 260 265 270gta ctt ctc agt tcg gcc tat ggt cat gtc aac tcg ccg atc gtc cag 864Val Leu Leu Ser Ser Ala Tyr Gly His Val Asn Ser Pro Ile Val Gln 275 280 285tac cga cac ttg ggc gcc ttc gcg gca gga acc caa tcc agc gcc agc 912Tyr Arg His Leu Gly Ala Phe Ala Ala Gly Thr Gln Ser Ser Ala Ser 290 295 300ccg act ctc tgc agc ccc gag cag gtc cgc gcc gga ggt tgc gtc aac 960Pro Thr Leu Cys Ser Pro Glu Gln Val Arg Ala Gly Gly Cys Val Asn305 310 315 320gtg ttc ggc gca ggc acg ccg agc ggc ttc tac gac ggt tcc agc gat 1008Val Phe Gly Ala Gly Thr Pro Ser Gly Phe Tyr Asp Gly Ser Ser Asp 325 330 335cgc ggt gaa cgc ttg cgc gtg gaa aac ttc ctg cag cag gcc cgc gcc 1056Arg Gly Glu Arg Leu Arg Val Glu Asn Phe Leu Gln Gln Ala Arg Ala 340 345 350gac tat gag gtc ggt ccg gtg acc ctg aca tcg atc agc gcc ttc acg 1104Asp Tyr Glu Val Gly Pro Val Thr Leu Thr Ser Ile Ser Ala Phe Thr 355 360 365cac agc aaa aag agc ggc ccc gac gac gcc gac ggg acg tct gac agt 1152His Ser Lys Lys Ser Gly Pro Asp Asp Ala Asp Gly Thr Ser Asp Ser 370 375 380ctg ctc cac gcg acc tac ggc gtt cgc tcc gac acc tgg acc caa gag 1200Leu Leu His Ala Thr Tyr Gly Val Arg Ser Asp Thr Trp Thr Gln Glu385 390 395 400ttc cgc gcc gcc tat tcc ggc cag cgc ctg cat tgg gtg gcg ggc gcc 1248Phe Arg Ala Ala Tyr Ser Gly Gln Arg Leu His Trp Val Ala Gly Ala 405 410 415tac tat ctc gac gag acc ctc aag caa aat cag cca ctt agc atc ttc 1296Tyr Tyr Leu Asp Glu Thr Leu Lys Gln Asn Gln Pro Leu Ser Ile Phe 420 425 430tac gat gga gat cgc ttc ggc ggc ctg ggc atc ccg gcc agg gcg gga 1344Tyr Asp Gly Asp Arg Phe Gly Gly Leu Gly Ile Pro Ala Arg Ala Gly 435 440 445gcc ttc gac ggc atc gcg caa aag agc tta agc caa aac act cag aaa 1392Ala Phe Asp Gly Ile Ala Gln Lys Ser Leu Ser Gln Asn Thr Gln Lys 450 455 460aca cgg tcg ata gcc gcc ttc ggc caa gcc gac tat acc ttg gac cgg 1440Thr Arg Ser Ile Ala Ala Phe Gly Gln Ala Asp Tyr Thr Leu Asp Arg465 470 475 480ttc acc ctg acc ttg ggc ggt cgt tac acc cat gaa cgc aag acg ttc 1488Phe Thr Leu Thr Leu Gly Gly Arg Tyr Thr His Glu Arg Lys Thr Phe 485 490 495gat cac ttc agc gcg acc cag gtc caa gca gga ggc ctt ggg aaa tac 1536Asp His Phe Ser Ala Thr Gln Val Gln Ala Gly Gly Leu Gly Lys Tyr 500 505 510ggt cct ctc ggc aag atc gtc tcg ctg agc gaa gcg ttc aag gct tcc 1584Gly Pro Leu Gly Lys Ile Val Ser Leu Ser Glu Ala Phe Lys Ala Ser 515 520 525gat ccg acc tgg cgc gcc gcg ctt tcc tac cgt ccc gcc gag cgt gtt 1632Asp Pro Thr Trp Arg Ala Ala Leu Ser Tyr Arg Pro Ala Glu Arg Val 530 535 540atg gtc tac ggc agc gtc gcc acc ggc ttt aag ggc ggc gcc ttc aac 1680Met Val Tyr Gly Ser Val Ala Thr Gly Phe Lys Gly Gly Ala Phe Asn545 550 555 560ggc ggg ttc ctg agc agc aac ccc aac aaa gcc ctc gcc gcg gtc aaa 1728Gly Gly Phe Leu Ser Ser Asn Pro Asn Lys Ala Leu Ala Ala Val Lys 565 570 575ccc gtc gca ccg gag aag gtg acc acc tac gaa ctg ggc ttc aag tcg 1776Pro Val Ala Pro Glu Lys Val Thr Thr Tyr Glu Leu Gly Phe Lys Ser 580 585 590agc ctg ttc gag cgt cgc ctg gtg gtc aac ggc gcg gct ttc tac aac 1824Ser Leu Phe Glu Arg Arg Leu Val Val Asn Gly Ala Ala Phe Tyr Asn 595 600 605agc tac gac aac gag cag atc ctg gcc aac acg gcc gtc gtc gtg gat 1872Ser Tyr Asp Asn Glu Gln Ile Leu Ala Asn Thr Ala Val Val Val Asp 610 615 620acc gtg acc ggc cct gtt acc gtg acg acg aac gtc ctg acc aac gcc 1920Thr Val Thr Gly Pro Val Thr Val Thr Thr Asn Val Leu Thr Asn Ala625 630 635 640cga aag gcc cac tcc cag ggc gtg gaa ttg gaa gta aag gcc gtc ccg 1968Arg Lys Ala His Ser Gln Gly Val Glu Leu Glu Val Lys Ala Val Pro 645 650 655atc ccg gat ctc gtc ctc agc ctg cag ccg gcc tgg ctg cga acg cgg 2016Ile Pro Asp Leu Val Leu Ser Leu Gln Pro Ala Trp Leu Arg Thr Arg 660 665 670ctg gac gag gcg ggc ttc tcc ggg gga acg tcg ctg gaa ggc aag caa 2064Leu Asp Glu Ala Gly Phe Ser Gly Gly Thr Ser Leu Glu Gly Lys Gln 675 680 685ctg gcc aat gcg ccg aag ttc tcg ctc tac gcc gcg gcg gac tac acc 2112Leu Ala Asn Ala Pro Lys Phe Ser Leu Tyr Ala Ala Ala Asp Tyr Thr 690 695 700ttc cat ctt gcc gac gac gac agc gtc aac gtc gcc ttc acc tcg gcc 2160Phe His Leu Ala Asp Asp Asp Ser Val Asn Val Ala Phe Thr Ser Ala705 710 715 720tac aag tcg cac cag ttc ttc gat tcg acg aac gcc ccc tat acc cag 2208Tyr Lys Ser His Gln Phe Phe Asp Ser Thr Asn Ala Pro Tyr Thr Gln 725 730 735cag gag ggc tac tgg gtg cac aac gcc agc ctg acc ttc aac tcc aga 2256Gln Glu Gly Tyr Trp Val His Asn Ala Ser Leu Thr Phe Asn Ser Arg 740 745 750aac cac tgg gat gtc ggg ttc aat gtc cga aac ctg acg ggc acg aag 2304Asn His Trp Asp Val Gly Phe Asn Val Arg Asn Leu Thr Gly Thr Lys 755 760 765tac tac aac tat ctg ttc gac gag ggg gcg acg ttc ggc ttc atc aac 2352Tyr Tyr Asn Tyr Leu Phe Asp Glu Gly Ala Thr Phe Gly Phe Ile Asn 770 775 780ggc gtc gtg gcc gcg ccg cgg acc tac agc gtg caa ttc aac ctg cat 2400Gly Val Val Ala Ala Pro Arg Thr Tyr Ser Val Gln Phe Asn Leu His785 790 795 800ctc 2403Leu 25 801 PRT Bacterium 2412.1 25Met Pro Arg Asp Thr Thr Pro Asn Phe Ala Pro Val Thr Thr Ala Lys 1 5 10 15Glu Gly Arg Arg His Arg Gly Ser Thr Ala Leu Arg Arg Leu Met Leu 20 25 30Thr Ala Ala Gly Ser Ala Leu Val Leu Gly Leu Ala Pro Lys Ala Leu 35 40 45Ala Gln Val Ala Val Pro Pro Ala Gly His Glu Ala Ser Gln Glu Val 50 55 60Gln Glu Ile Val Val Thr Ala Gln Arg Arg Ser Glu Asn Ile Gln Asn 65 70 75 80Val Pro Val Ser Val Gln Ala Leu Ser Ala Ala Gln Leu Glu Arg Glu 85 90 95Gly Ile Lys Gln Thr Ser Asp Ile Ala Arg Val Thr Pro Asn Val Thr 100 105 110Ile Ala Met Pro Asn Gly Glu Gly Asn Gln Pro Ala Val Thr Ile Arg 115 120 125Gly Ile Gly Leu Asn Asp Phe Asn Ser Asn Asn Ala Gly Pro Asn Ala 130 135 140Ile Tyr Val Asp Asp Val Tyr Ile Ser Ala Pro Ser Ala Gln Thr Phe145 150 155 160Gly Ile Phe Asp Ile Asn Gln Ile Gln Val Leu Lys Gly Pro Gln Gly 165 170 175Thr Leu Tyr Gly Arg Asn Ser Ser Gly Gly Ala Leu Val Phe Thr Ser 180 185 190Arg Ala Pro Ser Gln Asp Phe Ala Ala Asp Ala His Phe Asp Tyr Gly 195 200 205Ser Tyr Asn Thr Tyr Gln Leu Gln Ala Gly Val Gly Gly Pro Leu Ser 210 215 220Asp Gln Leu Ser Ala Arg Leu Ala Phe Val Val Asn His Ser Asp Gly225 230 235 240Phe Met His Asn Thr Leu Thr Gly Gly Ser Ala Ser Gly Thr Asp Asn 245 250 255Gln Ala Val Arg Leu Gln Leu Leu Tyr Arg Pro Asn Asp Arg Leu Lys 260 265 270Val Leu Leu Ser Ser Ala Tyr Gly His Val Asn Ser Pro Ile Val Gln 275 280 285Tyr Arg His Leu Gly Ala Phe Ala Ala Gly Thr Gln Ser Ser Ala Ser 290 295 300Pro Thr Leu Cys Ser Pro Glu Gln Val Arg Ala Gly Gly Cys Val Asn305 310 315 320Val Phe Gly Ala Gly Thr Pro Ser Gly Phe Tyr Asp Gly Ser Ser Asp 325 330 335Arg Gly Glu Arg Leu Arg Val Glu Asn Phe Leu Gln Gln Ala Arg Ala 340 345 350Asp Tyr Glu Val Gly Pro Val Thr Leu Thr Ser Ile Ser Ala Phe Thr 355 360 365His Ser Lys Lys Ser Gly Pro Asp Asp Ala Asp Gly Thr Ser Asp Ser 370 375 380Leu Leu His Ala Thr Tyr Gly Val Arg Ser Asp Thr Trp Thr Gln Glu385 390 395 400Phe Arg Ala Ala Tyr Ser Gly Gln Arg Leu His Trp Val Ala Gly Ala 405 410 415Tyr Tyr Leu Asp Glu Thr Leu Lys Gln Asn Gln Pro Leu Ser Ile Phe 420 425 430Tyr Asp Gly Asp Arg Phe Gly Gly Leu Gly Ile Pro Ala Arg Ala Gly 435 440 445Ala Phe Asp Gly Ile Ala Gln Lys Ser Leu Ser Gln Asn Thr Gln Lys 450 455 460Thr Arg Ser Ile Ala Ala Phe Gly Gln Ala Asp Tyr Thr Leu Asp Arg465 470 475 480Phe Thr Leu Thr Leu Gly Gly Arg Tyr Thr His Glu Arg Lys Thr Phe 485 490 495Asp His Phe Ser Ala Thr Gln Val Gln Ala Gly Gly Leu Gly Lys Tyr 500 505 510Gly Pro Leu Gly Lys Ile Val Ser Leu Ser Glu Ala Phe Lys Ala Ser 515 520 525Asp Pro Thr Trp Arg Ala Ala Leu Ser Tyr Arg Pro Ala Glu Arg Val 530 535 540Met Val Tyr Gly Ser Val Ala Thr Gly Phe Lys Gly Gly Ala Phe Asn545 550 555 560Gly Gly Phe Leu Ser Ser Asn Pro Asn Lys Ala Leu Ala Ala Val Lys 565 570 575Pro Val Ala Pro Glu Lys Val Thr Thr Tyr Glu Leu Gly Phe Lys Ser 580 585 590Ser Leu Phe Glu Arg Arg Leu Val Val Asn Gly Ala Ala Phe Tyr Asn 595 600 605Ser Tyr Asp Asn Glu Gln Ile Leu Ala Asn Thr Ala Val Val Val Asp 610 615 620Thr Val Thr Gly Pro Val Thr Val Thr Thr Asn Val Leu Thr Asn Ala625 630 635 640Arg Lys Ala His Ser Gln Gly Val Glu Leu Glu Val Lys Ala Val Pro 645 650 655Ile Pro Asp Leu Val Leu Ser Leu Gln Pro Ala Trp Leu Arg Thr Arg 660 665 670Leu Asp Glu Ala Gly Phe Ser Gly Gly Thr Ser Leu Glu Gly Lys Gln 675 680 685Leu Ala Asn Ala Pro Lys Phe Ser Leu Tyr Ala Ala Ala Asp Tyr Thr 690 695 700Phe His Leu Ala Asp Asp Asp Ser Val Asn Val Ala Phe Thr Ser Ala705 710 715 720Tyr Lys Ser His Gln Phe Phe Asp Ser Thr Asn Ala Pro Tyr Thr Gln 725 730 735Gln Glu Gly Tyr Trp Val His Asn Ala Ser Leu Thr Phe Asn Ser Arg 740 745 750Asn His Trp Asp Val Gly Phe Asn Val Arg Asn Leu Thr Gly Thr Lys 755 760 765Tyr Tyr Asn Tyr Leu Phe Asp Glu Gly Ala Thr Phe Gly Phe Ile Asn 770 775 780Gly Val Val Ala Ala Pro Arg Thr Tyr Ser Val Gln Phe Asn Leu His785 790 795 800Leu 26 1173 DNA Bacterium 2412.1 fccM 26atg gct cgc atc ggc ttc tct ttc gtc cca ccg ccg aca gct cgg ctc 48Met Ala Arg Ile Gly Phe Ser Phe Val Pro Pro Pro Thr Ala Arg Leu 1 5 10 15ggg aaa gtg act gtg tgg atg gaa ctg ggg cgt tcg tct gag cgc cgg 96Gly Lys Val Thr Val Trp Met Glu Leu Gly Arg Ser Ser Glu Arg Arg 20 25 30aga cga cgc aaa tcg agg cgg cgc gga ctc cag atc gag aac atc gcc 144Arg Arg Arg Lys Ser Arg Arg Arg Gly Leu Gln Ile Glu Asn Ile Ala 35 40 45tcg acc gtc gtc acg cca tcg tcg acg gcg tca ccc cgc ctt ggc gcc 192Ser Thr Val Val Thr Pro Ser Ser Thr Ala Ser Pro Arg Leu Gly Ala 50 55 60atc cgc gca agg ggg gat gga agg tcc ggg cga cac cac cgc cgc gcg 240Ile Arg Ala Arg Gly Asp Gly Arg Ser Gly Arg His His Arg Arg Ala 65 70 75 80gcc ctc ggg acc gaa tgt cat ggt cga ctt gcc cgt aga ata ggc agg 288Ala Leu Gly Thr Glu Cys His Gly Arg Leu Ala Arg Arg Ile Gly Arg 85 90 95cca ggt aag ggc gtc ccc ggc ggg gtc gcc att ctt ggc gaa ccg gac 336Pro Gly Lys Gly Val Pro Gly Gly Val Ala Ile Leu Gly Glu Pro Asp 100 105 110cca ggc gga gga cat cag ttg gcc cag cgc acg gtc ggc ggg cgt ggg 384Pro Gly Gly Gly His Gln Leu Ala Gln Arg Thr Val Gly Gly Arg Gly 115 120 125ccc ctc ggg cgg cca atc gaa cag acc caa ctc gtc gag ctt gaa cac 432Pro Leu Gly Arg Pro Ile Glu Gln Thr Gln Leu Val Glu Leu Glu His 130 135 140ccc gaa aac gta ggg aat ttc ggc tcc gtg ggt agc cgg cgc tct tcc 480Pro Glu Asn Val Gly Asn Phe Gly Ser Val Gly Ser Arg Arg Ser Ser145 150 155 160acc ctc ggt att acc gtt gaa ctg ata acg cca cac ggg cgc gcc ctg 528Thr Leu Gly Ile Thr Val Glu Leu Ile Thr Pro His Gly Arg Ala Leu 165 170 175gcg cac aag cgc ttc cga gaa ggc cga gac ccc ccg att gaa ctg att 576Ala His Lys Arg Phe Arg Glu Gly Arg Asp Pro Pro Ile Glu Leu Ile 180 185 190gtc gcc gaa gat gcg cgc gac cat ttc ctt ggg cgt ggc ccg gcc gtc 624Val Ala Glu Asp Ala Arg Asp His Phe Leu Gly Arg Gly Pro Ala Val 195 200 205gag ggg ata gca cgc cgc cac ggc ggc ggc ttg gtc gcc aaa ctg cgc 672Glu Gly Ile Ala Arg Arg His Gly Gly Gly Leu Val Ala Lys Leu Arg 210 215 220ctc cag ata ggc ttg gta gtc cgc tgg cgt ctc cat cgg cgc gcg ccc 720Leu Gln Ile Gly Leu Val Val Arg Trp Arg Leu His Arg Arg Ala Pro225 230 235 240gag gaa ggc gcg gcc ttc gtc ggc att ggt tcc gat cag gac ccg aac 768Glu Glu Gly Ala Ala Phe Val Gly Ile Gly Ser Asp Gln Asp Pro Asn 245 250 255cgg cgc cag ctg ccc cgc cgc gat cgc cgc gct gtc ggt ctg cgg cag 816Arg Arg Gln Leu Pro Arg Arg Asp Arg Arg Ala Val Gly Leu Arg Gln 260 265 270cac atg gcc atc gac gat cgg tcc ggt cgg acg cgg cct gcg cag gtc 864His Met Ala Ile Asp Asp Arg Ser Gly Arg Thr Arg Pro Ala Gln Val 275 280 285ccg cga tgc cgg gcg ggc cgc gtc ggc gcg cgc cat cag ggt ggc tgg 912Pro Arg Cys Arg Ala Gly Arg Val Gly Ala Arg His Gln Gly Gly Trp 290 295 300gtc ggt cga gcg cag tcg cga aag atc ggc gtc gag gcg ctc gcc cga 960Val Gly Arg Ala Gln Ser Arg Lys Ile Gly Val Glu Ala Leu Ala Arg305 310 315 320ggc ggc gct gtc ggc gag cgt cgc gag cgg tcg cgt cag ccc tgg act 1008Gly Gly Ala Val Gly Glu Arg Arg Glu Arg Ser Arg Gln Pro Trp Thr 325 330 335ttc gag gat agc gcc acg gaa gag acc ctt gct cag cgg cga ggt gag 1056Phe Glu Asp Ser Ala Thr Glu Glu Thr Leu Ala Gln Arg Arg Gly Glu 340 345 350cag aag tcc gat cgc gct cgc tcc ggc cga ttc acc aaa gac cgt cac 1104Gln Lys Ser Asp Arg Ala Arg Ser Gly Arg Phe Thr Lys Asp Arg His 355 360 365tcg gcc ggg gtc ccc tcc gaa ggc gcg ggc gtt gct ctg cac cca ccg 1152Ser Ala Gly Val Pro Ser Glu Gly Ala Gly Val Ala Leu His Pro Pro 370 375 380aag agc ggc gag aat gtc gag 1173Lys Ser Gly Glu Asn Val Glu385 390 27 391 PRT Bacterium 2412.1 27Met Ala Arg Ile Gly Phe Ser Phe Val Pro Pro Pro Thr Ala Arg Leu 1 5 10 15Gly Lys Val Thr Val Trp Met Glu Leu Gly Arg Ser Ser Glu Arg Arg 20 25 30Arg Arg Arg Lys Ser Arg Arg Arg Gly Leu Gln Ile Glu Asn Ile Ala 35 40 45Ser Thr Val Val Thr Pro Ser Ser Thr Ala Ser Pro Arg Leu Gly Ala 50 55 60Ile Arg Ala Arg Gly Asp Gly Arg Ser Gly Arg His His Arg Arg Ala 65 70 75 80Ala Leu Gly Thr Glu Cys His Gly Arg Leu Ala Arg Arg Ile Gly Arg 85 90 95Pro Gly Lys Gly Val Pro Gly Gly Val Ala Ile Leu Gly Glu Pro Asp 100 105 110Pro Gly Gly Gly His Gln Leu Ala Gln Arg Thr Val Gly Gly Arg Gly 115 120 125Pro Leu Gly Arg Pro Ile Glu Gln Thr Gln Leu Val Glu Leu Glu His 130 135 140Pro Glu Asn Val Gly Asn Phe Gly Ser Val Gly Ser Arg Arg Ser Ser145 150 155 160Thr Leu Gly Ile Thr Val Glu Leu Ile Thr Pro His Gly Arg Ala Leu 165 170 175Ala His Lys Arg Phe Arg Glu Gly Arg Asp Pro Pro Ile Glu Leu Ile 180 185 190Val Ala Glu Asp Ala Arg Asp His Phe Leu Gly Arg Gly Pro Ala Val 195 200 205Glu Gly Ile Ala Arg Arg His Gly Gly Gly Leu Val Ala Lys Leu Arg 210 215 220Leu Gln Ile Gly Leu Val Val Arg Trp Arg Leu His Arg Arg Ala Pro225 230 235 240Glu Glu Gly Ala Ala Phe Val Gly Ile Gly Ser Asp Gln Asp Pro Asn 245 250 255Arg Arg Gln Leu Pro Arg Arg Asp Arg Arg Ala Val Gly Leu Arg Gln 260 265 270His Met Ala Ile Asp Asp Arg Ser Gly Arg Thr Arg Pro Ala Gln Val 275 280 285Pro Arg Cys Arg Ala Gly Arg Val Gly Ala Arg His Gln Gly Gly Trp 290 295 300Val Gly Arg Ala Gln Ser Arg Lys Ile Gly Val Glu Ala Leu Ala Arg305 310 315 320Gly Gly Ala Val Gly Glu Arg Arg Glu Arg Ser Arg Gln Pro Trp Thr 325 330 335Phe Glu Asp Ser Ala Thr Glu Glu Thr Leu Ala Gln Arg Arg Gly Glu 340 345 350Gln Lys Ser Asp Arg Ala Arg Ser Gly Arg Phe Thr Lys Asp Arg His 355 360 365Ser Ala Gly Val Pro Ser Glu Gly Ala Gly Val Ala Leu His Pro Pro 370 375 380Lys Ser Gly Glu Asn Val Glu385 390 28 1179 DNA Bacterium 2412.1 fccN Citrate Utilization B 28atg cag cat aat gtc ctg gat ttc gtg acc aag acg cgc acg ggc gag 48Met Gln His Asn Val Leu Asp Phe Val Thr Lys Thr Arg Thr Gly Glu 1 5 10 15ccg cgc ccg gcc gaa acg ccc gcg atc atc gaa gcg cgc cgg acc atg 96Pro Arg Pro Ala Glu Thr Pro Ala Ile Ile Glu Ala Arg Arg Thr Met 20 25 30gag gtt tgc aac gcc tgt cgc tat tgc gaa ggc tac tgc gcg gtc ttt 144Glu Val Cys Asn Ala Cys Arg Tyr Cys Glu Gly Tyr Cys Ala Val Phe 35 40 45ccg gcc atg acc ctc aag cgg gag ttc gag gaa gcc gat ctc acc tac 192Pro Ala Met Thr Leu Lys Arg Glu Phe Glu Glu Ala Asp Leu Thr Tyr 50 55 60ctg gcc aat ctc tgt cac tcg tgc cgc ggc tgt tac tac gct tgc caa 240Leu Ala Asn Leu Cys His Ser Cys Arg Gly Cys Tyr Tyr Ala Cys Gln 65 70 75 80tac gcg ccg ccc cat gag ttc ggg atc aac gtg ccc aag gtg ctg gcc 288Tyr Ala Pro Pro His Glu Phe Gly Ile Asn Val Pro Lys Val Leu Ala 85 90 95gag gtc cgc acc gaa agc tac cag gcc cat gcc tgg ccg cag gcc gtc 336Glu Val Arg Thr Glu Ser Tyr Gln Ala His Ala Trp Pro Gln Ala Val 100 105 110gcc gtc gcc ttc gag cgt aac ggt ctg gtg gtg tcc ctg agc gct gca 384Ala Val Ala Phe Glu Arg Asn Gly Leu Val Val Ser Leu Ser Ala Ala 115 120 125ctc gcg atc gtt gtc gtg ctg ctg gga acg gcc ttc ttc aat gga tcg 432Leu Ala Ile Val Val Val Leu Leu Gly Thr Ala Phe Phe Asn Gly Ser 130 135 140gcg atg ttc cag gcg cac gcc tcg acg ccc ggc gca ggc ttc tac aag 480Ala Met Phe Gln Ala His Ala Ser Thr Pro Gly Ala Gly Phe Tyr Lys145 150 155 160gcc gtg ccc tat gcg gtc atg gtg agc gtc gcc ggc gcg atc ttc gcc 528Ala Val Pro Tyr Ala Val Met Val Ser Val Ala Gly Ala Ile Phe Ala 165 170 175tat gcc gcc ttg gcg atg ttc atc ggc ctt atc cgg ttt tgg aag acc 576Tyr Ala Ala Leu Ala Met Phe Ile Gly Leu Ile Arg Phe Trp Lys Thr 180 185 190gtg ggc ctt ggc ttg cgc gac gcc gtc gaa ccg cga acc ttg ttc cag 624Val Gly Leu Gly Leu Arg Asp Ala Val Glu Pro Arg Thr Leu Phe Gln 195 200 205gcg ctg aag gat gcg gcg acc ctg cgc tat ctc ggc ggg ggc ggc gat 672Ala Leu Lys Asp Ala Ala Thr Leu Arg Tyr Leu Gly Gly Gly Gly Asp 210 215 220ggc tgc aac gac gtc gac gct agc ttc tcg acc tca cgc cga cgt ttc 720Gly Cys Asn Asp Val Asp Ala Ser Phe Ser Thr Ser Arg Arg Arg Phe225 230 235 240cat cac gcc atg gcc tac ggc ttc ctg ctc tgt ttt gcc tcc acc tcc 768His His Ala Met Ala Tyr Gly Phe Leu Leu Cys Phe Ala Ser Thr Ser 245 250 255acc ggt acg gtc tac gac cac ctc ctg ggc tgg ccc gcg ccc tat ccc 816Thr Gly Thr Val Tyr Asp His Leu Leu Gly Trp Pro Ala Pro Tyr Pro 260 265 270ttc ttc agc ctg ccg gtg ctg ctg gga acg gtc ggc gga att ggg atc 864Phe Phe Ser Leu Pro Val Leu Leu Gly Thr Val Gly Gly Ile Gly Ile 275 280 285gtc atc ggc acg ctc gga ctg ctc tgg ctg aag ctg gtc ggc gac cag 912Val Ile Gly Thr Leu Gly Leu Leu Trp Leu Lys Leu Val Gly Asp Gln 290 295 300gag cct agg tcg aag gcg caa ttg ggc gcc gac acc gcg ctg ctg gtg 960Glu Pro Arg Ser Lys Ala Gln Leu Gly Ala Asp Thr Ala Leu Leu Val305 310 315 320ctg ctg ttc ctg atc agc gtg acg ggg ctg ttg ctg ctg gcg ctt cgg 1008Leu Leu Phe Leu Ile Ser Val Thr Gly Leu Leu Leu Leu Ala Leu Arg 325 330 335acg acg gcg gcc atg ggc gtg atc ctg acc gtg cac ctt gga ctg gtc 1056Thr Thr Ala Ala Met Gly Val Ile Leu Thr Val His Leu Gly Leu Val 340 345 350ttc tcg ttc ttc gcg acg atg ccg tac agc aag ttc gtg cac gga ctc 1104Phe Ser Phe Phe Ala Thr Met Pro Tyr Ser Lys Phe Val His Gly Leu 355 360 365tat cga acc gtc gcc ttg gtt cgt tac gcc gtc gag cgc aag gcg ctg 1152Tyr Arg Thr Val Ala Leu Val Arg Tyr Ala Val Glu Arg Lys Ala Leu 370 375 380gcc tcc ggg acg acg gag gaa gcg tct 1179Ala Ser Gly Thr Thr Glu Glu Ala Ser385 390 29 393 PRT Bacterium 2412.1 29Met Gln His Asn Val Leu Asp Phe Val Thr Lys Thr Arg Thr Gly Glu 1 5 10 15Pro Arg Pro Ala Glu Thr Pro Ala Ile Ile Glu Ala Arg Arg Thr Met 20 25 30Glu Val Cys Asn Ala Cys Arg Tyr Cys Glu Gly Tyr Cys Ala Val Phe 35 40 45Pro Ala Met Thr Leu Lys Arg Glu Phe Glu Glu Ala Asp Leu Thr Tyr 50 55 60Leu Ala Asn Leu Cys His Ser Cys Arg Gly Cys Tyr Tyr Ala Cys Gln 65 70 75 80Tyr Ala Pro Pro His Glu Phe Gly Ile Asn Val Pro Lys Val Leu Ala 85 90 95Glu Val Arg Thr Glu Ser Tyr Gln Ala His Ala Trp Pro Gln Ala Val 100 105 110Ala Val Ala Phe Glu Arg Asn Gly Leu Val Val Ser Leu Ser Ala Ala 115 120 125Leu Ala Ile Val Val Val Leu Leu Gly Thr Ala Phe Phe Asn Gly Ser 130 135 140Ala Met Phe Gln Ala His Ala Ser Thr Pro Gly Ala Gly Phe Tyr Lys145 150 155 160Ala Val Pro Tyr Ala Val Met Val Ser Val Ala Gly Ala Ile Phe Ala 165 170 175Tyr Ala Ala Leu Ala Met Phe Ile Gly Leu Ile Arg Phe Trp Lys Thr 180 185 190Val Gly Leu Gly Leu Arg Asp Ala Val Glu Pro Arg Thr Leu Phe Gln 195 200 205Ala Leu Lys Asp Ala Ala Thr Leu Arg Tyr Leu Gly Gly Gly Gly Asp 210 215 220Gly Cys Asn Asp Val Asp Ala Ser Phe Ser Thr Ser Arg Arg Arg Phe225 230 235 240His His Ala Met Ala Tyr Gly Phe Leu Leu Cys Phe Ala Ser Thr Ser 245 250 255Thr Gly Thr Val Tyr Asp His Leu Leu Gly Trp Pro Ala Pro Tyr Pro 260 265 270Phe Phe Ser Leu Pro Val Leu Leu Gly Thr Val Gly Gly Ile Gly Ile 275 280 285Val Ile Gly Thr Leu Gly Leu Leu Trp Leu Lys Leu Val Gly Asp Gln 290 295 300Glu Pro Arg Ser Lys Ala Gln Leu Gly Ala Asp Thr Ala Leu Leu Val305 310 315 320Leu Leu Phe Leu Ile Ser Val Thr Gly Leu Leu Leu Leu Ala Leu Arg 325 330 335Thr Thr Ala Ala Met Gly Val Ile Leu Thr Val His Leu Gly Leu Val 340 345 350Phe Ser Phe Phe Ala Thr Met Pro Tyr Ser Lys Phe Val His Gly Leu 355 360 365Tyr Arg Thr Val Ala Leu Val Arg Tyr Ala Val Glu Arg Lys Ala Leu 370 375 380Ala Ser Gly Thr Thr Glu Glu Ala Ser385 390 30 477 DNA Bacterium 2412.1 fccO Regulatory protein 30atg gcc gcg gcg aac acc ctg gga gcg act tcc ttg ggg tcg acg tca 48Met Ala Ala Ala Asn Thr Leu Gly Ala Thr Ser Leu Gly Ser Thr Ser 1 5 10 15tcc agg acg atc aga gcg tcg ttt ccg ccc aac tca agc gat ata cgt 96Ser Arg Thr Ile Arg Ala Ser Phe Pro Pro Asn Ser Ser Asp Ile Arg 20 25 30ttg agg cct tcg gcc gcg ccg gcc atg acc ttt ttt ccg gtc tgg gtc 144Leu Arg Pro Ser Ala Ala Pro Ala Met Thr Phe Phe Pro Val Trp Val 35 40 45gat ccg gtg aag ctg att ttg cga atg cca gga tgg cgg gtc att tcc 192Asp Pro Val Lys Leu Ile Leu Arg Met Pro Gly Trp Arg Val Ile Ser 50 55 60gcg ccg aga tcg tcg gcg tcg gtg atg atg tta atg acg ccc ggt ggg 240Ala Pro Arg Ser Ser Ala Ser Val Met Met Leu Met Thr Pro Gly Gly 65 70 75 80acg ata tcc ttg acc aag gcg cca aac cga agc gcc gtc aga ggc gtc 288Thr Ile Ser Leu Thr Lys Ala Pro Asn Arg Ser Ala Val Arg Gly Val 85 90 95gtc gcc gcc ggc ttg agg atg acc gtg ttg ccg gcc agc agg gcc gcc 336Val Ala Ala Gly Leu Arg Met Thr Val Leu Pro Ala Ser Arg Ala Ala 100 105 110ggg atc ttg aac gcc atc aac agc atc ggg aaa ttc cag ggg acg atg 384Gly Ile Leu Asn Ala Ile Asn Ser Ile Gly Lys Phe Gln Gly Thr Met 115 120 125cag ccc acc acg cct agg ggg cgt cta tgc acc tct acg cgg ccc gtc 432Gln Pro Thr Thr Pro Arg Gly Arg Leu Cys Thr Ser Thr Arg Pro Val 130 135 140gcg tcg tct ctg acc acg cga ggc ggc aga tcg agc gag gtg aag 477Ala Ser Ser Leu Thr Thr Arg Gly Gly Arg Ser Ser Glu Val Lys145 150 155 31 159 PRT Bacterium 2412.1 31Met Ala Ala Ala Asn Thr Leu Gly Ala Thr Ser Leu Gly Ser Thr Ser 1 5 10 15Ser Arg Thr Ile Arg Ala Ser Phe Pro Pro Asn Ser Ser Asp Ile Arg 20 25 30Leu Arg Pro Ser Ala Ala Pro Ala Met Thr Phe Phe Pro Val Trp Val 35 40 45Asp Pro Val Lys Leu Ile Leu Arg Met Pro Gly Trp Arg Val Ile Ser 50 55 60Ala Pro Arg Ser Ser Ala Ser Val Met Met Leu Met Thr Pro Gly Gly 65 70 75 80Thr Ile Ser Leu Thr Lys Ala Pro Asn Arg Ser Ala Val Arg Gly Val 85 90 95Val Ala Ala Gly Leu Arg Met Thr Val Leu Pro Ala Ser Arg Ala Ala 100 105 110Gly Ile Leu Asn Ala Ile Asn Ser Ile Gly Lys Phe Gln Gly Thr Met 115 120 125Gln Pro Thr Thr Pro Arg Gly Arg Leu Cys Thr Ser Thr Arg Pro Val 130 135 140Ala Ser Ser Leu Thr Thr Arg Gly Gly Arg Ser Ser Glu Val Lys145 150 155 32 1056 DNA Bacterium 2412.1 fccP 32atg gcg gcg cct cct tct gcg cgc cta gag atg cag gtt gaa ttg cac 48Met Ala Ala Pro Pro Ser Ala Arg Leu Glu Met Gln Val Glu Leu His 1 5 10 15gct gta ggt ccg cgg cgc ggc cac gac gcc gtt gat gaa gcc gaa cgt 96Ala Val Gly Pro Arg Arg Gly His Asp Ala Val Asp Glu Ala Glu Arg 20 25 30cgc ccc ctc gtc gaa cag ata gtt gta gta ctt cgt gcc cgt cag gtt 144Arg Pro Leu Val Glu Gln Ile Val Val Val Leu Arg Ala Arg Gln Val 35 40 45tcg gac att gaa ccc gac atc cca gtg gtt tct gga gtt gaa ggt cag 192Ser Asp Ile Glu Pro Asp Ile Pro Val Val Ser Gly Val Glu Gly Gln 50 55 60gct ggc gtt gtg cac cca gta gcc ctc ctg ctg ggt ata ggg ggc gtt 240Ala Gly Val Val His Pro Val Ala Leu Leu Leu Gly Ile Gly Gly Val 65 70 75 80cgt cga atc gaa gaa ctg gtg cga ctt gta ggc cga ggt gaa ggc gac 288Arg Arg Ile Glu Glu Leu Val Arg Leu Val Gly Arg Gly Glu Gly Asp 85 90 95gtt gac gct gtc gtc gtc ggc aag atg gaa ggt gta gtc cgc cgc ggc 336Val Asp Ala Val Val Val Gly Lys Met Glu Gly Val Val Arg Arg Gly 100 105 110gta gag cga gaa ctt cgg cgc att ggc cag ttg ctt gcc ttc cag cga 384Val Glu Arg Glu Leu Arg Arg Ile Gly Gln Leu Leu Ala Phe Gln Arg 115 120 125cgt tcc ccc gga gaa gcc cgc ctc gtc cag ccg cgt tcg cag cca ggc 432Arg Ser Pro Gly Glu Ala Arg Leu Val Gln Pro Arg Ser Gln Pro Gly 130 135 140cgg ctg cag gct gag gac gag atc cgg gat cgg gac ggc ctt tac ttc 480Arg Leu Gln Ala Glu Asp Glu Ile Arg Asp Arg Asp Gly Leu Tyr Phe145 150 155 160caa ttc cac gcc ctg gga gtg ggc ctt tcg ggc gtt ggt cag gac gtt 528Gln Phe His Ala Leu Gly Val Gly Leu Ser Gly Val Gly Gln Asp Val 165 170 175cgt cgt cac ggt aac agg gcc ggt cac ggt atc cac gac gac ggc cgt 576Arg Arg His Gly Asn Arg Ala Gly His Gly Ile His Asp Asp Gly Arg 180 185 190gtt ggc cag gat ctg ctc gtt gtc gta gct gtt gta gaa agc cgc gcc 624Val Gly Gln Asp Leu Leu Val Val Val Ala Val Val Glu Ser Arg Ala 195 200 205gtt gac cac cag gcg acg ctc gaa cag gct cga ctt gaa gcc cag ttc 672Val Asp His Gln Ala Thr Leu Glu Gln Ala Arg Leu Glu Ala Gln Phe 210 215 220gta ggt ggt cac ctt ctc cgg tgc gac ggg ttt gac cgc ggc gag ggc 720Val Gly Gly His Leu Leu Arg Cys Asp Gly Phe Asp Arg Gly Glu Gly225 230 235 240ttt gtt ggg gtt gct gct cag gaa ccc gcc gtt gaa ggc gcc gcc ctt 768Phe Val Gly Val Ala Ala Gln Glu Pro Ala Val Glu Gly Ala Ala Leu 245 250 255aaa gcc ggt ggc gac gct gcc gta gac cat aac acg ctc ggc ggg acg 816Lys Ala Gly Gly Asp Ala Ala Val Asp His Asn Thr Leu Gly Gly Thr 260 265 270gta gga aag cgc ggc gcg cca ggt cgg atc gga agc ctt gaa cgc ttc 864Val Gly Lys Arg Gly Ala Pro Gly Arg Ile Gly Ser Leu Glu Arg Phe 275 280 285gct cag cga gac gat ctt gcc gag agg acc gta ttt ccc aag gcc tcc 912Ala Gln Arg Asp Asp Leu Ala Glu Arg Thr Val Phe Pro Lys Ala Ser 290 295 300tgc ttg gac ctg ggt cgc gct gaa gtg atc gaa cgt ctt gcg ttc atg 960Cys Leu Asp Leu Gly Arg Ala Glu Val Ile Glu Arg Leu Ala Phe Met305 310 315 320ggt gta acg acc gcc caa ggt cag ggt gaa ccg gtc caa ggt ata gtc 1008Gly Val Thr Thr Ala Gln Gly Gln Gly Glu Pro Val Gln Gly Ile Val 325 330 335ggc ttg gcc gaa ggc ggc tat cga ccg tgt ttt ctg agt gtt ttg gct 1056Gly Leu Ala Glu Gly Gly Tyr Arg Pro Cys Phe Leu Ser Val Leu Ala 340 345 350 33 352 PRT Bacterium 2412.1 33Met Ala Ala Pro Pro Ser Ala Arg Leu Glu Met Gln Val Glu Leu His 1 5 10 15Ala Val Gly Pro Arg Arg Gly His Asp Ala Val Asp Glu Ala Glu Arg 20 25 30Arg Pro Leu Val Glu Gln Ile Val Val Val Leu Arg Ala Arg Gln Val 35 40 45Ser Asp Ile Glu Pro Asp Ile Pro Val Val Ser Gly Val Glu Gly Gln 50 55 60Ala Gly Val Val His Pro Val Ala Leu Leu Leu Gly Ile Gly Gly Val 65 70 75 80Arg Arg Ile Glu Glu Leu Val Arg Leu Val Gly Arg Gly Glu Gly Asp 85 90 95Val Asp Ala Val Val Val Gly Lys Met Glu Gly Val Val Arg Arg Gly 100 105 110Val Glu Arg Glu Leu Arg Arg Ile Gly Gln Leu Leu Ala Phe Gln Arg 115 120 125Arg Ser Pro Gly Glu Ala Arg Leu Val Gln Pro Arg Ser Gln Pro Gly 130 135 140Arg Leu Gln Ala Glu Asp Glu Ile Arg Asp Arg Asp Gly Leu Tyr Phe145 150 155 160Gln Phe His Ala Leu Gly Val Gly Leu Ser Gly Val Gly Gln Asp Val 165 170 175Arg Arg His Gly Asn Arg Ala Gly His Gly Ile His Asp Asp Gly Arg 180 185 190Val Gly Gln Asp Leu Leu Val Val Val Ala Val Val Glu Ser Arg Ala 195 200 205Val Asp His Gln Ala Thr Leu Glu Gln Ala Arg Leu Glu Ala Gln Phe 210 215 220Val Gly Gly His Leu Leu Arg Cys Asp Gly Phe Asp Arg Gly Glu Gly225 230 235 240Phe Val Gly Val Ala Ala Gln Glu Pro Ala Val Glu Gly Ala Ala Leu 245 250 255Lys Ala Gly Gly Asp Ala Ala Val Asp His Asn Thr Leu Gly Gly Thr 260 265 270Val Gly Lys Arg Gly Ala Pro Gly Arg Ile Gly Ser Leu Glu Arg Phe 275 280 285Ala Gln Arg Asp Asp Leu Ala Glu Arg Thr Val Phe Pro Lys Ala Ser 290 295 300Cys Leu Asp Leu Gly Arg Ala Glu Val Ile Glu Arg Leu Ala Phe Met305 310 315 320Gly Val Thr Thr Ala Gln Gly Gln Gly Glu Pro Val Gln Gly Ile Val 325 330 335Gly Leu Ala Glu Gly Gly Tyr Arg Pro Cys Phe Leu Ser Val Leu Ala 340 345 350 34 477 DNA Bacterium 2412.1 fccQ Leucine Regulatory Protein Homolog 34atg ctg agg tta cgc cac atg atg cca gcg ctg gac gcg atc gac cgc 48Met Leu Arg Leu Arg His Met Met Pro Ala Leu Asp Ala Ile Asp Arg 1 5 10 15aag atc att ggc ctg ctt cgg gtc aat ggc cgc atg ccc aac aat gag 96Lys Ile Ile Gly Leu Leu Arg Val Asn Gly Arg Met Pro Asn Asn Glu 20 25 30ttg gcg cag aag gta ggg ctt tcg cct tcc gcc tgt ctg cga cgc gtc 144Leu Ala Gln Lys Val Gly Leu Ser Pro Ser Ala Cys Leu Arg Arg Val 35 40 45aag ttg ctc gag tcg aac ggg gtg atc cga ggg tat tgt gca ttg gtt 192Lys Leu Leu Glu Ser Asn Gly Val Ile Arg Gly Tyr Cys Ala Leu Val 50 55 60gcc gag cag tcg ttg gac gcc agc gtg gtg gcg atc gtc cgg ata acc 240Ala Glu Gln Ser Leu Asp Ala Ser Val Val Ala Ile Val Arg Ile Thr 65 70 75 80ttg gac aag cag acc gag gac tat ctg aat cgg ttc gag gag gcc gtt 288Leu Asp Lys Gln Thr Glu Asp Tyr Leu Asn Arg Phe Glu Glu Ala Val 85 90 95cgg cgg cat ccc gag atc gct gag tgc ttt ctg atg acc ggc gac gca 336Arg Arg His Pro Glu Ile Ala Glu Cys Phe Leu Met Thr Gly Asp Ala 100 105 110gac tac atc ctt cgg gct acc gcg ccg agc acg gcc gcc tac gag caa 384Asp Tyr Ile Leu Arg Ala Thr Ala Pro Ser Thr Ala Ala Tyr Glu Gln 115 120 125atc cac aag gaa gtc ctt tct cgg ctt ccc ggg gtg gcg cgc atc cat 432Ile His Lys Glu Val Leu Ser Arg Leu Pro Gly Val Ala Arg Ile His 130 135 140tcg agc ttc gcc atc cgc agc gtg ctg tcg tcg gtc gca agg ccc 477Ser Ser Phe Ala Ile Arg Ser Val Leu Ser Ser Val Ala Arg Pro145 150 155 35 159 PRT Bacterium 2412.1 35Met Leu Arg Leu Arg His Met Met Pro Ala Leu Asp Ala Ile Asp Arg 1 5 10 15Lys Ile Ile Gly Leu Leu Arg Val Asn Gly Arg Met Pro Asn Asn Glu 20 25 30Leu Ala Gln Lys Val Gly Leu Ser Pro Ser Ala Cys Leu Arg Arg Val 35 40 45Lys Leu Leu Glu Ser Asn Gly Val Ile Arg Gly Tyr Cys Ala Leu Val 50 55 60Ala Glu Gln Ser Leu Asp Ala Ser Val Val Ala Ile Val Arg Ile Thr 65 70 75 80Leu Asp Lys Gln Thr Glu Asp Tyr Leu Asn Arg Phe Glu Glu Ala Val 85 90 95Arg Arg His Pro Glu Ile Ala Glu Cys Phe Leu Met Thr Gly Asp Ala 100 105 110Asp Tyr Ile Leu Arg Ala Thr Ala Pro Ser Thr Ala Ala Tyr Glu Gln 115 120 125Ile His Lys Glu Val Leu Ser Arg Leu Pro Gly Val Ala Arg Ile His 130 135 140Ser Ser Phe Ala Ile Arg Ser Val Leu Ser Ser Val Ala Arg Pro145 150 155 36 1074 DNA Bacterium 2412.1 fccR 36atg gcg gct gac gag atc tcc cag ctc agg aga acc cat ccc cac cgt 48Met Ala Ala Asp Glu Ile Ser Gln Leu Arg Arg Thr His Pro His Arg 1 5 10 15ctt gat gac tcc ctg gag gcg gtc gct gat ctg cag gta gcc ctc cgc 96Leu Asp Asp Ser Leu Glu Ala Val Ala Asp Leu Gln Val Ala Leu Arg 20 25 30gtc aat ccg gcc gaa atc ctg cgt atg aag gct tcc tcc gcg cca aag 144Val Asn Pro Ala Glu Ile Leu Arg Met Lys Ala Ser Ser Ala Pro Lys 35 40 45ctc gcg cga ggc ggc cag atc gcg atg gta ccc agc cgt cag cca agg 192Leu Ala Arg Gly Gly Gln Ile Ala Met Val Pro Ser Arg Gln Pro Arg 50 55 60cgc acg caa cac gat ctc gcc cgc gga gag gcc gtc gcg ggg cac atc 240Arg Thr Gln His Asp Leu Ala Arg Gly Glu Ala Val Ala Gly His Ile 65 70 75 80cgc cat gga ctc atc cac gac ccg gag gtc gac cag cgc gat agc ttg 288Arg His Gly Leu Ile His Asp Pro Glu Val Asp Gln Arg Asp Ser Leu 85 90 95tcc gac ctt ggt gcg aat ggc gac ctc ctc ctg agc tgg agc cat tgc 336Ser Asp Leu Gly Ala Asn Gly Asp Leu Leu Leu Ser Trp Ser His Cys 100 105 110ttc ggt cct aat gcg cgt gaa gct gac cag cgg ccc ggt ttc gga cat 384Phe Gly Pro Asn Ala Arg Glu Ala Asp Gln Arg Pro Gly Phe Gly His 115 120 125tcc gta tcc ggt ggt cag gac gat gcc gcg ccc cgc ggc ctg aag agc 432Ser Val Ser Gly Gly Gln Asp Asp Ala Ala Pro Arg Gly Leu Lys Ser 130 135 140cag gcc gcg ggg cag cgc cgc tcc gcc gac gag cac ctg cca tcc ggt 480Gln Ala Ala Gly Gln Arg Arg Ser Ala Asp Glu His Leu Pro Ser Gly145 150 155 160gaa atc cgc cgt ctg gcc cga agg cga att gag cat cat ctg caa aag 528Glu Ile Arg Arg Leu Ala Arg Arg Arg Ile Glu His His Leu Gln Lys 165 170 175tgt ggg cac gca atg aga gaa ggt cac ctt ctc cgc gcg ctg aag ctc 576Cys Gly His Ala Met Arg Glu Gly His Leu Leu Arg Ala Leu Lys Leu 180 185 190cac cag ttg ctc ggg aac ata gcg ccc tgg gta gac ttg ctt gca gcc 624His Gln Leu Leu Gly Asn Ile Ala Pro Trp Val Asp Leu Leu Ala Ala 195 200 205gac cat cgt cgc cac gaa ggg cat tcc cca cgc atg agc atg gaa cat 672Asp His Arg Arg His Glu Gly His Ser Pro Arg Met Ser Met Glu His 210 215 220cgg ggt gat cgg cat gta gac cgt atc gcg ccc cag cct ggc gta acc 720Arg Gly Asp Arg His Val Asp Arg Ile Ala Pro Gln Pro Gly Val Thr225 230 235 240gtc tcc aag ggc cag cgt cgc cat cac cgc cag ggt gtg cag cac cag 768Val Ser Lys Gly Gln Arg Arg His His Arg Gln Gly Val Gln His Gln 245 250 255ctg acg atg cga ata gta gac gcc ctt cgg acg ccc ggt cgt gcc gct 816Leu Thr Met Arg Ile Val Asp Ala Leu Arg Thr Pro Gly Arg Ala Ala 260 265 270ggt gta gaa ggt cgt cgc ccg cgt gtt ctc gtc gaa gtc cgg gaa gtc 864Gly Val Glu Gly Arg Arg Pro Arg Val Leu Val Glu Val Arg Glu Val 275 280 285gaa ccg agg gcg cgc ggc cgc cat cag ggc ctc ata ctc ccc cag cac 912Glu Pro Arg Ala Arg Gly Arg His Gln Gly Leu Ile Leu Pro Gln His 290 295 300cca ggg atg ggc ggc ttc cgc ctc gtc gtc ctt cag atg gac gac ccc 960Pro Gly Met Gly Gly Phe Arg Leu Val Val Leu Gln Met Asp Asp Pro305 310 315 320gcg cag gct ggg caa ttg gtc gat cac ctc ctc gag gat cgg cag gaa 1008Ala Gln Ala Gly Gln Leu Val Asp His Leu Leu Glu Asp Arg Gln Glu 325 330 335atc ggt gtg gct cag cgc cag ggt ggc gcc cgc atg ctc gag cgt gta 1056Ile Gly Val Ala Gln Arg Gln Gly Gly Ala Arg Met Leu Glu Arg Val 340 345 350gcg cag gtc gtc tcg ggc 1074Ala Gln Val Val Ser Gly 355 37 358 PRT Bacterium 2412.1 37Met Ala Ala Asp Glu Ile Ser Gln Leu Arg Arg Thr His Pro His Arg 1 5 10 15Leu Asp Asp Ser Leu Glu Ala Val Ala Asp Leu Gln Val Ala Leu Arg 20 25 30Val Asn Pro Ala Glu Ile Leu Arg Met Lys Ala Ser Ser Ala Pro Lys 35 40 45Leu Ala Arg Gly Gly Gln Ile Ala Met Val Pro Ser Arg Gln Pro Arg 50 55 60Arg Thr Gln His Asp Leu Ala Arg Gly Glu Ala Val Ala Gly His Ile 65 70 75 80Arg His Gly Leu Ile His Asp Pro Glu Val Asp Gln Arg Asp Ser Leu 85 90 95Ser Asp Leu Gly Ala Asn Gly Asp Leu Leu Leu Ser Trp Ser His Cys 100 105 110Phe Gly Pro Asn Ala Arg Glu Ala Asp Gln Arg Pro Gly Phe Gly His 115 120 125Ser Val Ser Gly Gly Gln Asp Asp Ala Ala Pro Arg Gly Leu Lys Ser 130 135 140Gln Ala Ala Gly Gln Arg Arg Ser Ala Asp Glu His Leu Pro Ser Gly145 150 155 160Glu Ile Arg Arg Leu Ala Arg Arg Arg Ile Glu His His Leu Gln Lys 165 170 175Cys Gly His Ala Met Arg Glu Gly His Leu Leu Arg Ala Leu Lys Leu 180 185 190His Gln Leu Leu Gly Asn Ile Ala Pro Trp Val Asp Leu Leu Ala Ala 195 200 205Asp His Arg Arg His Glu Gly His Ser Pro Arg Met Ser Met Glu His 210 215 220Arg Gly Asp Arg His Val Asp Arg Ile Ala Pro Gln Pro Gly Val Thr225 230 235 240Val Ser Lys Gly Gln Arg Arg His His Arg Gln Gly Val Gln His Gln 245 250 255Leu Thr Met Arg Ile Val Asp Ala Leu Arg Thr Pro Gly Arg Ala Ala 260 265 270Gly Val Glu Gly Arg Arg Pro Arg Val Leu Val Glu Val Arg Glu Val 275 280 285Glu Pro Arg Ala Arg Gly Arg His Gln Gly Leu Ile Leu Pro Gln His 290 295 300Pro Gly Met Gly Gly Phe Arg Leu Val Val Leu Gln Met Asp Asp Pro305 310 315 320Ala Gln Ala Gly Gln Leu Val Asp His Leu Leu Glu Asp Arg Gln Glu 325 330 335Ile Gly Val Ala Gln Arg Gln Gly Gly Ala Arg Met Leu Glu Arg Val 340 345 350Ala Gln Val Val Ser Gly 355 38 552 DNA Bacterium 2412.1 fccS 38atg cag gcg ctg gcc gga ata ggc ggc gcg gaa ctc ttg ggt cca ggt 48Met Gln Ala Leu Ala Gly Ile Gly Gly Ala Glu Leu Leu Gly Pro Gly 1 5 10 15gtc gga gcg aac gcc gta ggt cgc gtg gag cag act gtc aga cgt ccc 96Val Gly Ala Asn Ala Val Gly Arg Val Glu Gln Thr Val Arg Arg Pro 20 25 30gtc ggc gtc gtc ggg gcc gct ctt ttt gct gtg cgt gaa ggc gct gat 144Val Gly Val Val Gly Ala Ala Leu Phe Ala Val Arg Glu Gly Ala Asp 35 40 45cga tgt cag ggt cac cgg acc gac ctc ata gtc ggc gcg ggc ctg ctg 192Arg Cys Gln Gly His Arg Thr Asp Leu Ile Val Gly Ala Gly Leu Leu 50 55 60cag gaa gtt ttc cac gcg caa gcg ttc acc gcg atc gct gga acc gtc 240Gln Glu Val Phe His Ala Gln Ala Phe Thr Ala Ile Ala Gly Thr Val 65 70 75 80gta gaa gcc gct cgg cgt gcc tgc gcc gaa cac gtt gac gca acc tcc 288Val Glu Ala Ala Arg Arg Ala Cys Ala Glu His Val Asp Ala Thr Ser 85 90 95ggc gcg gac ctg ctc ggg gct gca gag agt cgg gct ggc gct gga ttg 336Gly Ala Asp Leu Leu Gly Ala Ala Glu Ser Arg Ala Gly Ala Gly Leu 100 105 110ggt tcc tgc cgc gaa ggc gcc caa gtg tcg gta ctg gac gat cgg cga 384Gly Ser Cys Arg Glu Gly Ala Gln Val Ser Val Leu Asp Asp Arg Arg 115 120 125gtt gac atg acc ata ggc cga act gag aag tac ttt cag cct gtc att 432Val Asp Met Thr Ile Gly Arg Thr Glu Lys Tyr Phe Gln Pro Val Ile 130 135 140agg tcg gta gag cag ttg cag cct gac ggc ctg att gtc cgt gcc cga 480Arg Ser Val Glu Gln Leu Gln Pro Asp Gly Leu Ile Val Arg Ala Arg145 150 155 160cgc cga acc gcc cgt cag cgt gtt gtg cat gaa ccc gtc gga gtg gtt 528Arg Arg Thr Ala Arg Gln Arg Val Val His Glu Pro Val Gly Val Val 165 170 175gac gac gaa ggc cag gcg ggc gct 552Asp Asp Glu Gly Gln Ala Gly Ala 180 39 184 PRT Bacterium 2412.1 39Met Gln Ala Leu Ala Gly Ile Gly Gly Ala Glu Leu Leu Gly Pro Gly 1 5 10 15Val Gly Ala Asn Ala Val Gly Arg Val Glu Gln Thr Val Arg Arg Pro 20 25 30Val Gly Val Val Gly Ala Ala Leu Phe Ala Val Arg Glu Gly Ala Asp 35 40 45Arg Cys Gln Gly His Arg Thr Asp Leu Ile Val Gly Ala Gly Leu Leu 50 55 60Gln Glu Val Phe His Ala Gln Ala Phe Thr Ala Ile Ala Gly Thr Val 65 70 75 80Val Glu Ala Ala Arg Arg Ala Cys Ala Glu His Val Asp Ala Thr Ser 85 90 95Gly Ala Asp Leu Leu Gly Ala Ala Glu Ser Arg Ala Gly Ala Gly Leu 100 105 110Gly Ser Cys Arg Glu Gly Ala Gln Val Ser Val Leu Asp Asp Arg Arg 115 120 125Val Asp Met Thr Ile Gly Arg Thr Glu Lys Tyr Phe Gln Pro Val Ile 130 135 140Arg Ser Val Glu Gln Leu Gln Pro Asp Gly Leu Ile Val Arg Ala Arg145 150 155 160Arg Arg Thr Ala Arg Gln Arg Val Val His Glu Pro Val Gly Val Val 165 170 175Asp Asp Glu Gly Gln Ala Gly Ala 180 40 531 DNA Bacterium 2412.1 fccT 40atg ggc gtc cgc ggc gaa gtc ttg gct cgg cgc tct gga cgt gaa cac 48Met Gly Val Arg Gly Glu Val Leu Ala Arg Arg Ser Gly Arg Glu His 1 5 10 15caa ggc ccc acc gct gga gtt gcg ccc ata gag cgt acc ttg cgg tcc 96Gln Gly Pro Thr Ala Gly Val Ala Pro Ile Glu Arg Thr Leu Arg Ser 20 25 30ttt gag aac ctg gat ctg gtt gat gtc gaa gat tcc gaa ggt ctg ggc 144Phe Glu Asn Leu Asp Leu Val Asp Val Glu Asp Ser Glu Gly Leu Gly 35 40 45cga cgg ggc gct gat ata gac atc gtc gac ata gat cgc gtt cgg gcc 192Arg Arg Gly Ala Asp Ile Asp Ile Val Asp Ile Asp Arg Val Arg Ala 50 55 60ggc gtt gtt gga att gaa gtc gtt gag gcc gat gcc gcg gat cgt cac 240Gly Val Val Gly Ile Glu Val Val Glu Ala Asp Ala Ala Asp Arg His 65 70 75 80cgc cgg ctg gtt gcc ttc gcc gtt ggg cat ggc gat ggt gac gtt ggg 288Arg Arg Leu Val Ala Phe Ala Val Gly His Gly Asp Gly Asp Val Gly 85 90 95cgt cac tcg ggc gat atc gct ggt ctg ttt gat ccc ttc gcg ctc gag 336Arg His Ser Gly Asp Ile Ala Gly Leu Phe Asp Pro Phe Ala Leu Glu 100 105 110ctg cgc tgc cga cag cgc ctg cac cga gac cgg cac att ctg aat gtt 384Leu Arg Cys Arg Gln Arg Leu His Arg Asp Arg His Ile Leu Asn Val 115 120 125ctc gct gcg gcg ctg cgc ggt gac gac gat ctc ctg cac ctc ctg cga 432Leu Ala Ala Ala Leu Arg Gly Asp Asp Asp Leu Leu His Leu Leu Arg 130 135 140cgc ctc gtg acc agc cgg cgg aac cgc cac ctg cgc gag cgc ctt ggg 480Arg Leu Val Thr Ser Arg Arg Asn Arg His Leu Arg Glu Arg Leu Gly145 150 155 160cgc aag acc cag cac cag ggc gct gcc ggc cgc cgt cag cat gag cct 528Arg Lys Thr Gln His Gln Gly Ala Ala Gly Arg Arg Gln His Glu Pro 165 170 175tcg 531Ser 41 177 PRT Bacterium 2412.1 41Met Gly Val Arg Gly Glu Val Leu Ala Arg Arg Ser Gly Arg Glu His 1 5 10 15Gln Gly Pro Thr Ala Gly Val Ala Pro Ile Glu Arg Thr Leu Arg Ser 20 25 30Phe Glu Asn Leu Asp Leu Val Asp Val Glu Asp Ser Glu Gly Leu Gly 35 40 45Arg Arg Gly Ala Asp Ile Asp Ile Val Asp Ile Asp Arg Val Arg Ala 50 55 60Gly Val Val Gly Ile Glu Val Val Glu Ala Asp Ala Ala Asp Arg His 65 70 75 80Arg Arg Leu Val Ala Phe Ala Val Gly His Gly Asp Gly Asp Val Gly 85 90 95Arg His Ser Gly Asp Ile Ala Gly Leu Phe Asp Pro Phe Ala Leu Glu 100 105 110Leu Arg Cys Arg Gln Arg Leu His Arg Asp Arg His Ile Leu Asn Val 115 120 125Leu Ala Ala Ala Leu Arg Gly Asp Asp Asp Leu Leu His Leu Leu Arg 130 135 140Arg Leu Val Thr Ser Arg Arg Asn Arg His Leu Arg Glu Arg Leu Gly145 150 155 160Arg Lys Thr Gln His Gln Gly Ala Ala Gly Arg Arg Gln His Glu Pro 165 170 175Ser 42 26 DNA Artificial Sequence Description of Artificial Sequence synthetic oligonucleotide 42gttgcgatgg tcgcgagaat aagcgt 26 43 25 DNA Artificial Sequence Description of Artificial Sequence synthetic oligonucleotide 43agtaggccgt agttgcccga agttc 25 44 28 DNA Artificial Sequence Description of Artificial Sequence synthetic nucleotide 44acccgattat cgtcaatgac gaacgagg 28 45 24 DNA Artificial Sequence Description of Artificial Sequence synthetic oligonucleotide 45accatggtcg cgtagtcgtc tctc 24 46 23 DNA Artificial Sequence Description of Artificial Sequence synthetic oligonucleotide 46gatccaacgc ttggagggac tgg 23 47 25 DNA Artificial Sequence Description of Artificial Sequence synthetic oligonucleotide 47gaccattcga tgacgtcgct ttgcg 25 48 26 DNA Artificial Sequence Description of Artificial Sequence synthetic oligonucleotide 48aattgttcga tccgatcacg gccacg 26 49 24 DNA Artificial Sequence Description of Artificial Sequence synthetic oligonucleotide 49tgagacccac cgtcgtgcta tcgt 24 50 24 DNA Artificial Sequence Description of Artificial Sequence synthetic oligonucleotide 50acgcgaccat atcccgctcg tgat 24 51 27 DNA Artificial Sequence Description of Artificial Sequence synthetic oligonucleotide 51cctggtcaac gacgttgagc agacatt 27 52 25 DNA Artificial Sequence Description of Artificial Sequence synthetic oligonucleotide 52gacgaggatt tcgagggcct aaagg 25 53 23 DNA Artificial Sequence Description of Artificial Sequence synthetic oligonucleotide 53tcgatctcga ccagcgggaa ctc 23 54 640 DNA Bacterium 2412.1 54gttgcgatgg tcgcgagaat aagcgtgcga agtgggagga tgtgaagatg ggggccagga 60gtatgtgtgc gggacggttc ggacgcttct gcattggctt ggcttcatcg gttgccgtga 120ctctaggggg agcctccgcc gccggcgcgg caaccgcgac ggattttccg gtccgcagga 180ccgatctggg ccaggttcag ggactggccg gggacgtgat gagctttcgc ggaataccct 240atgcagcgcc gccggtgggc gggctgcgtt ggaagccgcc ccaacacgcc cggccctggg 300cgggcgttcg ccccgccacc caatttggct ccgactgctt cggcgcggcc tatcttcgca 360aaggcagcct cgcccccggc gtgagcgagg actgtcttta cctcaacgta tgggcgccgt 420caggcgctaa acccggccag taccccgtca tggtctgggt ctacggcggc ggcttcgccg 480gcggcacggc cgccatgccc tactacgacg gcgaggcgct tgcgcgacag ggcgtcgtcg 540tggtgacgtt taactatcgg acgaacatcc tgggcttttt cgcccatcct ggtctctcgc 600gcgagagccc caccggaact tcgggcaact acggcctact 640 55 475 DNA Bacterium 2412.1 55acccgattat cgtcaatgac gaacgaggcc gccaacggac cggcgctgaa gcccaccaac 60gcgagagggg cgagcagtac ggcggcggtc ctgctgggct ccaaggcgat cacgtccgcc 120accagaaacg gctgcagcgc cagccagaac aggccaaagc cacaagcgct tgcgatgagc 180gcgacgggcg tgccggcgtg aagcaggccg atgacaagac cggcctgcag cactgcgccg 240gcggccagaa ccgtacgggc gtgcacgcgc gcaccgagcc aggatgctgc aagagcaccc 300gccacctgga aggccaggct gcccgcgatc gcggcgccga ccgtggccgg ggcgaaatgg 360tgttgcgcgg ccaggcgctc caggtagttc catgccgccc cgatgccggc gttttgaaga 420aacgccgcga gcgccacgac catcagggcc ggagagacga ctacgcgacc atggt 475 56 447 DNA Bacterium 2412.1 56gatccaacgc ttggagggac tggtcggcgg cgctctcttc gaccgaacca gccggaccat 60gaccgagacc gcgcttggca aggagctgct gccggtggcc cgccgaacgc ttgagtttct 120ggacaattcg ctgttcgcct cgcccaagct gcgcgaaccg cgctggaccg acatcagcat 180tttttgcgtg cagaccgccg cgttccgcgt tctgccgcgc gcggcccggc gcttcatgga 240tgaaaatccc cgactgcgcc tgaggatcat cgatgttccg gctgtcgaag gcgcggaact 300ggtggcgcga ggggaagcgg agttcggtat cagcatcgag agcctgcttc cgtccggcct 360gcgtttcgag gctcttcacg aggacccgtt tggcttggcg tgccatcgga gccatcgcct 420ggcgcaaagc gacgtcatcg aatggtc 447 57 536 DNA Bacterium 2412.1 57tgagacccag ccgttcgtgc ttatcgtcgg cggcggtcaa ggcggtctag cgcttggcgc 60gcgcctccgt cagctccagg tcccgactct gatcgtcgat cagcacccac gggtggggga 120ccaatggcga tcgcggtacg catcgctctg cctgcacgat ccagtctggt acgaccacct 180tccttacctg ccgtttcccg atacttggcc ggtttatacg cccaaggaca agatcggcga 240ttggctcgaa gcttatgcgc aggcgatgga gctgctggtc tggtgttcga ccagatgcgt 300gtccgccgtc tatgacgccg aagccgggcg atggaccgtc accctgcgcc gaggcgagga 360gaccagcgtc atccgccccg cgcatctggt cctggcgacg ggcaacgccg gcaagccgcg 420cgttccgcgc ttcaagggcc aagcgcagtt cgaaggtccg atcctgcact cgagcgccta 480tcggagcggg gctgatttca aaggacggcg cgtggccgtg atcggatcga acaatt 536 58 715 DNA Bacterium 2412.1 58acgcgaccat atcccgctcg tgatcggctg cgggcccgtg ggcatggcgg tcatcgccgc 60gctccggggt ctgggcgtcg gaccgatcat cgcggccgac ttcaatccgg cgcgtcggag 120cctggcggcg cgcatgggcg ccgatattgt catcgacccg gcggagcggt ccccctacga 180cgaatggcgg gataccgcgg cggcgtcagg cctggccgga ctggcggggg cgccagcgtc 240gctgcggacc tgtctggtct tcgagtgtgt cggcctgcca ggaatgctgc gtcagatcat 300ggaaggcgcc ccggcggagt cggagatcat cgtcgtcggg gcctgcatgg agcccgatag 360cctcgagccg atgatggcga tgcataaggc tctgacgctg aattttcgcg aacctacacg 420atcgaggagt tcgccgaggt ccttcggatg atcggtgagg gcgagctcca cgtcgagccg 480ttgctcagcc aacccatcgg cctggaagac cttccggggg tcttcgacaa agcgcccggg 540agggccgggg gcgccaaggt cctcgtcgac ccctggcgct gacgccgccg aaaaccaacg 600acagaaaaca acgggcggga ggaacaattg ggtagcgatg agcaagacga tcctctcgtc 660ggcggcggtc gtcagttgct tctcaagcaa tgtctgctca acgtcgttga ccagg 715 59 686 DNA Bacterium 2412.1 59tcgatctcga ccagcgggaa ctcgccggcg aaccggccga cggaaaccga cgtcagccgg 60ttggtgtcca tcagtgcgcc gagtgcggtc gggaggaacg cgttgtcgcg gaagatcgtg 120aaggcgttcg cgctgccgac aaactggttg acgaaggcgc cgaggttggt gtggctatag 180gcataggtgc cttccgcata gagcttgacg cgttctgagg cctcgaactc gccgcggagg 240aagccattgt agcgccgctg gtccggagcg aagccgagat tgacgcgcgg accgtcgccg 300ccgctctgga aggaactgct ggtaaagctt ccgtagttaa aggtcgcaag cgtaccgccg 360ggaagaaagg tgacgccctt cagcggaccc gatgtgatca ggccgccgta ggcgccgcgc 420gagcttcgga tgtcgggaac caccgtgacg cccgtcgggg cgccgggcac gggatattgc 480cccgccgcgc gatcatacca cgcccgatcg gtggcctgat cggcgcgaat gccgtcctcg 540tgatagtatt cgaccgcggc aaggagatgt gcgcgccctt gggcaaacga cttgccggcc 600gccagcgatc cgcccaccga ggccagatcg ttgcggctcg agacgccggt ctggacgttc 660gcctttaggc cctcgaaatc ctcgtc 686

Claims

1. An isolated nucleic acid molecule comprising a nucleotide sequence that encodes a fumonisin degradative enzyme, wherein said nucleotide sequence is selected from the group consisting of:a) a nucleotide sequence that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO: 7; and b) the nucleotide sequence set forth in SEQ ID NO:6.

2. An expression cassette comprising a nucleic acid molecule of claim 1, wherein said nucleic acid molecule is operably linked to a promoter that drives expression in a plant cell.

3. The expression cassette of claim 2, wherein said promoter is a constitutive promoter.

4. An expression cassette comprising a nucleic acid molecule of claim 1, wherein said nucleic acid molecule is operably linked to a promoter that drives expression in a microorganism.

5. The expression cassette of claim 4, wherein said promoter is a constitutive promoter.

6. A vector for delivery of a nucleic acid molecule to a host cell, wherein said vector comprises a nucleic acid molecule of claim 1.

7. A host cell containing the vector of claim 6.

8. A host cell comprising in its genome at least one nucleic acid molecule of claim 1.

9. The host cell of claim 8, wherein said host cell is a cell of a plant selected from the group consisting of maize, sorghum, wheat, tomato, soybean, alfalfa, sunflower, Brassica, cotton, and rice.

10. A transformed plant cell having incorporated into its genome at least one nucleic acid molecule of claim 1.

11. A transformed plant having incorporated into its genome at least one nucleic acid molecule of claim 1.

12. Transformed seed of the plant of claim 11.

13. A plant having stably incorporated within its genome a nucleotide sequence encoding a fumonisin degradative enzyme, wherein said nucleotide sequence is selected from the group consisting of:a) a nucleotide sequence that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO: 7; and b) the nucleotide sequence set forth in SEQ ID NO:6.

14. A plant capable of degrading fumonisin, said plant having stably incorporated within its genome at least one nucleotide sequence encoding a fumonisin degradative enzyme from Bacterium 2412.1 operably linked to a plant promoter, wherein said nucleotide sequence is selected from the group consisting of:a) a nucleotide sequence that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO: 7; and b) the nucleotide sequence set forth in SEQ ID NO:6.

15. The plant of claim 14, wherein said plant is selected from the group consisting of maize, sorghum, wheat, tomato, soybean, alfalfa, sunflower, Brassica, cotton, and rice.

16. A method of making an enzyme useful in degrading fumonisin, said method comprising:a) expressing a nucleic acid molecule of claim 1 in a recombinantly engineered cell; and b) purifying said enzyme.

17. A method of making an enzyme useful in degrading fumonisin, said method comprising:a) expressing a nucleic acid molecule of claim 1 in a plant; and b) purfying said enzyme from the plant seed or other plant parts.

18. A genetically engineered ruminal microorganism comprising at least one nucleic acid molecule of claim 1.

19. A probiotic composition comprising the genetically engineered ruminal microorganism of claim 18.

20. A feed inoculant composition comprising the genetically engineered ruminal microorganism of claim 18.