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 fumonisin or AP1. 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 33rd 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 postharvest 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 to visible mold to 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, U.S. Pat. No. 6,025,188, and U.S. Pat. No. 6,229,071, 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 enzymes capable of degrading AP1 are necessary for the further detoxification of fumonisin.

SUMMARY OF THE INVENTION

Compositions and methods for catabolism and detoxification of fumonisin and fumonisin-degradation products as well as fumonisin-related toxins 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 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 are steps in a catabolic pathway for fumonisin. Thus, organisms can be genetically modified to provide for the catabolism and detoxification of fumonisin and fumonisin-related toxins.

In particular, expression cassettes for expression of the enzymes 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 Exophiala spinifera.

FIG. 2 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

The catabolic pathway for detoxification and degradation of fumonisin is provided. Particularly, enzymes involved in the degradation of fumonisin from Exophiala spinifera (American Type Culture Collection Deposit No. 74269) and nucleotide sequences encoding such enzymes are disclosed. Such enzymes and nucleotide sequences find use in the breakdown of fumonisin and fumonisin-related toxins as well as degradation products thereof. 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 Exophiala spinifera is provided in FIG. 1 . The present invention encompasses enzymes and nucleotide sequences encoding the enzymes involved in this degradation pathway for fumonisin. Compositions of the invention include a flavin monooxygenase, an aldehyde dehydrogenase, a permease, and a p-glycoprotein that are involved in the fumonisin degradation pathway. In particular, the present invention provides for isolated nucleic acid molecules comprising nucleotide sequences encoding the amino acid sequences shown in SEQ ID NOS:3, 5, 8, and 11, or the nucleotide sequences encoding the DNA sequences obtained from the overlapping clones deposited in a bacterial host with the American Type Culture Collection and assigned Accession Number PTA-299. By “DNA sequence obtained from the overlapping clones” is intended that the DNA sequence of the fumonisin degrading enzymes can be obtained by sequencing the individual clones which together comprise the entire fumonisin degrading enzymes. 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:1, 2, 4, 6, 7, 9 and 10, the DNA sequences obtained from the overlapping clones deposited in a bacterial host with the American Type Culture Collection and assigned Accession Number PTA-299, and fragments and variants thereof.

Ten plasmids containing overlapping clones were deposited with the American Type Culture Collection, Manassas, Va., and assigned Accession Number PTA-299. The plasmids designated as F_perm3.5 and F_perm4.4 contain common sequences at the regions were they overlap to form the nucleotide sequence encoding a permease. The plasmids designated as F_p-glyco1L4, F_p-glyco5.13, and F_glyco6.43 contain common sequences at the regions were they overlap to form the nucleotide sequence encoding a p-glycoprotein. And the plasmids designated F_Alde1.1, F_Alde2.2, and F_Alde2.5 contain common sequences at the regions were they overlap to form the nucleotide sequence of an aldehyde dehydrogenase. One of skill in the art by sequencing the clones and aligning the overlap may obtain the entire sequence of the permease, the p-glycoprotein, and the aldehyde dehydrogenase.

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.

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.

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 fragment proteins 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, 25, 30, 50, 100, 150, 200, or 250, 300, 400, 500, 600, 700, 800, 900, 1,000, 1,100, 1,200 contiguous amino acids, or up to the total number of amino acids present in a full-length fumonisin-degrading protein of the invention (for example, 545, 487, 525, 1,263 amino acids for SEQ ID NOS:3, 5, 8 and 11, respectively). 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 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 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 16, 20, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 800, 900, 1,000, 1,100, 1,200, 1,300, or 1,400, 1500, 1,600, 1,800, 2,000, 2,200, 2,400, 2,600, 2,800, 3,000, 3,200, 3,400, 3,600, 3,800, 3,900 nucleotides, or up to the number of nucleotides present in a full-length fumonisin-degrading nucleotide sequence disclosed herein (for example, 1,691, 1,638, 1,464, 1,764, 1,578, 3,999, 3,792 nucleotides for SEQ ID NOS:1, 2, 4, 6, 7, 9, and 10 respectively).

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. (1983) 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 or catabolize fumonisin. Obviously, the mutations that will be made in the DNA encoding the variant must not place the sequence out of reading frame and preferably will not create complementary regions that could produce secondary mRNA structure. See, EP Patent 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 coding sequences can be manipulated to create a new fumonisin-catabolizing 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-catabolizing 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.

The carboxylesterase and amine oxidase have been previously described in U.S. Pat. No. 5,716,820, U.S. Pat. No. 6,025,188, and U.S. Pat. No. 6,229,071. Such disclosures are herein incorporated by reference. Thus, the sequences of the invention can be used in combination with those previously disclosed or disclosed in U.S. Pat. No. 6,211,435 and U.S. Pat. No. 6,211,434, both entitled “Amino Polyolamine Oxidase Polynucleotides and Related Polypeptides and Methods of Use”, herein incorporated by reference. These exemplary amino polyolamine oxidase nucleotide sequences are set forth in SEQ ID NOs: 16, 18, 20, 22, 24, 26, 28, 30, and 32. SEQ ID NO:16 is the same as SEQ ID NO:5 from U.S. Pat. No. 6,211,435; SEQ ID NO:18 is the same as SEQ ID NO:10 from U.S. Pat. No. 6,211,435; and SEQ ID NO:20 is the same as SEQ ID NO:22 from U.S. Pat. No. 6,211,435. The nucleotide sequences set forth in SEQ ID NOs: 16, 18, and 20 encode polypeptides having the amino acid sequences set forth in SEQ ID NOs: 17, 19, and 21, respectively (which are the same as SEQ ID NOs: 6, 11, and 23 from U.S. Pat. No. 6,211,435, respectively). Amino polyolamine oxidase nucleotide sequences of U.S. Pat. No. 6,211,434, with introns removed, are set forth in SEQ ID NOs:22 (SEQ ID NO:35 from U.S. Pat. No. 6,211,434), 24 (SEQ ID NO:37 from U.S. Pat. No. 6,211,434), 26 (SEQ ID NO:39 from U.S. Pat. No. 6,211,434), 28 (SEQ ID NO:41 from U.S. Pat. No. 6,211,434), 30 (SEQ ID NO:43 from U.S. Pat. No. 6,211,434), and 32 (SEQ ID NO:45 from U.S. Pat. No. 6,211,434). The nucleotide sequences set forth in SEQ ID NOs: 22, 24, 26, 28, 30, and 32 encode polypeptides having the amino acid sequences set forth in SEQ ID NOs: 23, 25, 27, 29, 31, and 33, respectively (which are the same as SEQ ID NOs: 36, 38, 40, 42, 44, and 46 from U.S. Pat. No. 6,211,434, respectively). The enzymes and nucleotide sequences of the present invention provide a means for continued catabolism of the fumonisin-degradation products obtained after degradation with at least the carboxylesterase and amine oxidase.

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, N.Y.); 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 ester linkage 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 on 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 linkage in fumonisin. Two examples of such enzymes are ESP1 and BEST1 found in U.S. Pat. No. 5,716,820, U.S. Pat. No. 6,025,188, and U.S. Pat. No. 6,229,071. The ESP1 nucleotide sequence is set forth in SEQ ID NO:12 and is the same as SEQ ID NO:15 from U.S. Pat. No. 6,025,188. This nucleotide sequence encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO:13 (which is the same as SEQ ID NO:10 from U.S. Pat. No. 6,025,188). The BEST1 nucleotide sequence is set forth in SEQ ID NO:14 and is the same as SEQ ID NO:11 from U.S. Pat. No. 6,025,188. This nucleotide sequence encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO: 15 (which is the same as SEQ ID NO:12 from U.S. Pat. No. 6,025,188).

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.

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 (nonrecombinant) 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 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 sequences 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 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, the entire fumonisin-degrading sequences 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 organism or as a diagnostic assay to determine the presence of coding sequences in 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 Tm 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-degradative protein and hybridize to the fumonisin-degrading sequences 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). Alignments are performed using the default parameters of the above mentioned programs. 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 metabolize fumonisin and resist its toxic effects.

Fumonisin is produced in the intercellular spaces (apoplast) of Fusarium-infected maize cells. Thus, the apoplast is the preferred location for esterase and deaminase, flavin amine oxidase 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 and reroute the fumonisin or degradation products 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. Such a pump useful in the invention and disclosed herein is a P-glycoprotein homolog.

More complete catabolism of fumonisin in transgenic organisms may be provided by esterase and deaminase enzymes. Exophiala enzymes that can further oxidize fumonisin breakdown-products are not detected extracellularly. Such enzymes in all likelihood exist in the cytoplasm, where adequate cofactors such as NAD + or NADP are found. The fumonisin-induced metabolite transporter is predicted to provide transport of degradation products into cells where they can be further broken down by other enzymes. In this manner, a permease enzyme may be utilized in a heterologous system to transport either AP1 precursors or fumonisin degradation products into the cytoplasm.

The monooxygenase is expected to result in the oxidation of 2-OP to a compound that lacks a keto group, having instead a terminal aldehyde group, or possibly a carboxylate group. See, for example, Trudgill et al. (1984) in Microbial Degradation of Organic Compounds, ed. Gibson (Microbiology Series Vol. 13, Marcel Dekker, New York), Chapter 6; and Davey and Trudgill (1977) Eur. J. Biochem. 74:115.

This reaction 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. The metabolism of trans-cyclohexane-1,2 diol by Acinetobacter provides a model for the activity of a Baeyer-Villiger monooxygenase on 2-OP. This diol is first oxidized to ortho hydroxy cyclohexanone and then a monooxygen is inserted between the quinone 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 it is predicted oxygen is inserted between carbons 2 and 3 followed by spontaneous cleavage to a C22 aldehyde and acetic acid. Further oxidation by an aldehyde dehydrogenase would convert this compound to a carboxylic acid; other catabolic products would also be possible given the high reactivity of the aldehyde group. Additional steps include the use of an aldehyde dehydrogenase to result in the oxidation of the aldehyde product of fumonisin to a hydroxy carboxylic acid.

It is recognized that the DNA sequences of the invention 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.

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 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. 1 7:7891-7903; 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. See, for example, Campbell and Gowri (1990) Plant Physiol. 92:1-11 for a discussion of host-preferred codon usage. 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, N.Y.), 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.

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 on plants (epiphytes) to deliver enzymes to potential target crops. Epiphytes can be gram-positive or gram-negative bacteria, 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.

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.

In this manner, at least one of the genes encoding a 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 pesticide, 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, Spirillum; 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, 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, 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 nucleotide 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 Exophiala spinifera (American Type Culture Collection Accession No. 74269), during the processing of grain for animal or human food consumption, during the processing of plant material for silage, or in 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 Exophiala spinifera (American Type Culture Collection Accession No. 74269), 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 enzymes or microorganisms 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, R. C. (1990) Principles of Cereal Science and Technology, American Assn. of Cereal Chemists, Inc. (especially Chapters 5, 6 and 7); Jones, J. M. (1992) Food Safety, Eagan Press, St. Paul, Minn. (especially Chapters 7 and 9); and Jelen, P. (1985) Introduction to Food Processing, Restan Publ. Co., 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 from Exophiala spinifera (American Type Culture Collection Accession No. 74269), showed a range of activity from about pH 3 to about pH 6, and the esterase from the bacterium of the American Type Culture Collection Accession No. 55552 showed a range of activity from about pH 6 to about pH 9 (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 6 to pH 9.

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 enzymes from Exophiala spinifera, American Type Culture Collection Accession No. 74269, showed a range of activity for esterase 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 6 to pH 9.

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 ruminal 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 degradative enzymes of Exophiala spinifera (American Type Culture Collection Accession No. 74269).

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

Fungal and Bacterial Isolates

Exophiala isolates from maize were isolated as described in U.S. Pat. No. 5,716,820, U.S. Pat. No. 6,025,188, and U.S. Pat. No. 6,229,071, herein incorporated by reference.

Isolation Methods

Direct isolation of black yeasts from seed was accomplished by plating 100 microliters of seed wash fluid onto YPD or Sabouraud agar augmented with cycloheximide (500 mg/liter) and chloramphenicol (50 mg/liter). Plates were incubated at room temperature for 7-14 days, and individual pigmented colonies that arose were counted and cultured for analysis of fumonisin-degrading ability as described above.

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, 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.

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.

Enzyme Activity of Culture Filtrate and Mycelium

Exophiala spinifera isolate 2141.10 was grown on YPD agar for 1 week, and conidia were harvested, suspended in sterile water, and used at 105 conidia per ml to inoculate sterile Fries mineral salts medium containing 1 mg/ml purified FB1 (Sigma Chemical Co.). After 2 weeks incubation at 28° C. in the dark, cultures were filtered through 0.45 micron cellulose acetate filters and rinsed with Fries mineral salts. Fungal mycelium was suspended in 15 mL of 0.1% FB1, pH 5.2+1 mM EDTA+3 μg/mL Pepstatin A+1.5 μg/mL Leupeptin and disrupted in a Bead Beater™ using 0.1 mm beads and one minute pulses, with ice cooling. Hyphal pieces were collected by filtering through Spin X™ (0.22 μm), and both mycelial supernatant and original culture filtrates were assayed for fumonisin modification by methods outlined above.

Preparation of Crude Culture Filtrate

Agar cultures grown as above were used to inoculate YPD broth cultures (500 ml) in conical flasks at a final concentration of 10 5 conidia per ml culture. Cultures were incubated 5 days at 28° C. without agitation and mycelia harvested by filtration through 0.45 micron filters under vacuum. The filtrate was discarded, and the mycelial mat was washed and resuspended in sterile carbon-free, mineral salts medium (1 g/liter NH 3 NO 4 ; 1 g/liter NaH 2 PO 4 ; 0.5 g/liter MgCl 2 ; 0.1 g/liter NaCl; 0.13 g/liter CaCl 2 ; 0.02 g/liter FeSO 4 .7H 2 O, pH 4.5) containing 0.5 mg/ml alkaline hydrolyzed crude FB1. After 3-5 days at 28° C. in the dark with no agitation the cultures were filtered through low protein binding 0.45 micron filters to recover the culture filtrate. Phenylmethyl sulfonyl fluoride (PMSF) was added to a concentration of 2.5 mM and the culture filtrate was concentrated using an Amicon™ YM10 membrane in a stirred cell at room temperature and resuspended in 50 mM sodium acetate, pH 5.2 containing 10mM CaCl 2 . The crude culture filtrate (approx. 200-fold concentrated) was stored at −20° C.

To obtain preparative amounts of enzyme-hydrolyzed fumonisin, 10 mg of FB1 (Sigma) was dissolved in 20 mL of 50 mM sodium acetate at pH 5.2+10 mM CaCl 2 , and 0.25 mL of 200×concentrated crude culture filtrate of 2141.10 was added. The solution was incubated at 37° C. for 14 hours, and then cooled to room temperature. The reaction mixture was brought to approximately pH 9.5 by addition of 0.4 mL of 4 N KOH, and the mixture was extracted twice with 10 mL ethyl acetate. The combined organic layers were dried under LN 2 and resuspended in dH 2 O. 2.5 milligrams of organic extracted material were analyzed by Fast Atom Bombardment (FAB) mass spectrometry. The resulting mass spectrum showed a major ion at M/z (+1)=406 mass units, indicating the major product of enzymatic hydrolysis was AP1, which has a calculated molecular weight of 405.

EXAMPLE 2

Preparation of AP1-induced and Non-induced Mycelium

Liquid cultures of Exophiala spinifera isolate 2141.10 were prepared from YPD agar plates (Yeast Extract 10 gm, Bacto-Peptone 20 gm, Dextrose 0.5 gm, Bacto-Agar 15 gm per liter of water). Aliquots (400-500 μL) of a water suspension of E. spinifera cells from YPD agar were spread uniformly onto 150×15 mm YPD agar plates with 4 mm sterile glass beads. The plates were incubated at room temperature for 6-7 days. The mycelia/conidia were transferred from the agar plates into Mineral Salts Medium (MSM) (Na 2 HPO 4 .7H 2 O 0.2 gm, NH 4 Cl 1.0 gm, CaCl 2 .2H 2 O 0.01 gm, FeSO 4 .7H 2 O 0.02 gm per liter of distilled water, pH 4.5) and centrifuged at 5000 × g, 4° C., 20 minutes to pellet the cells. The cell pellet was rinsed once in 40 mL MSM and recentrifuged. The rinsed cell pellet was used to inoculate MSM at a 1:19 ratio of packed cells: MSM. The culture was supplemented with AP1 to a final concentration of 0.5-1.0 mg/ml and incubated at 28° C., 100 rpm, in the dark to induce catabolic enzymes. The supernatants were removed by filtration through 0.45 cellulose acetate. The remaining mycelial mat was washed with sterile MSM and then frozen in liquid nitrogen for storage.

EXAMPLE 3

Effect of FB1 and AP1 on Maize Coleoptiles

Maize coleoptiles from 4 day dark-grown germinated maize seeds were excised above the growing point and placed in 96-well microliter plates in the presence of 60 microliters of sterile distilled water containing FB1 or AP1 at approximately equimolar concentrations of 1.5, 0.5, 0.15, 0.05, 0.015, 0.005, 0.0015, or 0.0005 millimolar, along with water controls. After 2 days in the dark at 28° C. the coleoptiles were placed in the light and incubated another 3 days. Injury or lack thereof was evaluated as follows:

	0	.0005	.0015	.005	.015	.05	.15	.5	1.5	mM
FB1	−	−	−	−	+/−	+	+	+	+
AP1	−	−	−	−	−	−	−	−	+
+ = brown necrotic discoloration of coleoptile
− = no symptoms (same as water control)

The results (see table above) indicate there is at least a 30-fold difference in toxicity between FB1 and AP1 to maize coleoptiles of this genotype. This is in general agreement with other studies where the toxicity of the two compounds was compared for plant tissues. In Lemna tissues, AP1 was approximately 40-fold less toxic (Vesonder et al. (1992) Arch. Environ. Contam. Toxicol. 23:464-467 (1992)). Studies with both AAL toxin and FB1 in tomato also indicate the hydrolyzed version of the molecule is much less toxic (Gilchrist et al. (1992) Mycopathologia 117: 57-64). Lamprecht et al. also observed an approximate 100-fold reduction in toxicity to tomato by AP1 versus FB1 (Lamprecht et al. (1994) Phytopathology 84:383-391).

EXAMPLE 4

Effect of FBI and AP1 on Maize Tissue Cultured Cells

Black Mexican Sweet, BMS

FB1 or AP1 at various concentrations was added to suspensions of BMS cells growing in liquid culture medium in 96-well polystyrene plates. After 1 week the cell density in wells was observed under low power magnification and growth of toxin-treated wells was compared to control wells that received water. Growth of BMS cells was significantly inhibited at 0.4 micromolar FB1, but no inhibition was observed until 40 micromolar AP1. This represents an approximate 100-fold difference in toxicity to maize tissue-cultured cells. Similarly Van Asch et al. observed significant inhibition of maize callus grown on solid medium at 1.4 micromolar FB1 (Van Asch et al. (1992) Phytopathology 82:1330-1332). AP1 was not tested in that study, however.

EXAMPLE 5

The polynucleotides were identified using a proprietary transcript imaging method that compares transcript patterns in two samples and allows cloning of differentially expressed fragments. This technology was developed by CuraGen® (New Haven, Conn.) (see PCT patent application Ser. No. WO 97/1 5690, published May 1, 1997 and claiming priority from U.S. application Ser. No. 08/663,823, which issued as U.S. Pat. No. 5,972,693, all of which are hereby incorporated by reference). Fluorescently-tagged, PCR amplified cDNA fragments representing expressed transcripts can be visualized as bands or peaks on a gel tracing, and the cDNA from differentially expressed (induced or suppressed) bands can be recovered from a duplicate gel, cloned, and sequenced. Known cDNAs can be identified without the need for cloning, by matching the predicted size and partially known sequence of specific bands on the tracing.

Two RNA samples were obtained from cultures of E. spinifera grown for a specified period in a mineral salts medium containing either AP1 (induced condition) or gamma-aminobutyric acid (ABA; non-induced condition) as a sole carbon source. In the induced condition, fumonisin esterase, amine oxidase, enzyme activities are detected, whereas in the non-induced condition these activities are not detected. The methods used for induction of and detection of enzyme activity are described earlier (see Example 2 and Example 5). RNA was extracted from induced mycelium by Tri-Reagent methods (Molecular Research Center Inc., Cincinnati, Ohio) only using frozen tissue samples ground with a mortar and pestle 2-fold and up to 79-fold and greater until slushy and adding an additional extraction after the phase separation by extracting the aqueous phase one time with phenol, and two times with a phenol:chloroform:isoamyl alcohol mixture. The RNAs were submitted for CuraGen® transcript imaging to detect cDNA fragments that are induced specifically in the presence AP1. In the resulting gel tracing several bands were found which showed induction of at least 10-fold in AP1-grown cells as compared to cells grown in ABA. One set of induced fragments can be matched to the fumonisin esterase cDNA. The cloned bands and possible functions are provided in Table 2. Highly induced bands and their likely function are provided in Tables 2 and 3.

TABLE 2
	Best BLAST	BLAST Hit Name,
Clone ID	Hit	source, size	Prob	from-to	Function
Monooxygenase
M1a0-388	A28550	cyclohexanone monooxygenase, Acinetobacter	1.4e−22	339-414	Baeyer-Villiger oxidation of
		(flavin monooxygenase or FMO)EC 1.14.13.22			2-OP1 (AP1-N1), utilizing
		Length = 543			molecular oxygen and
					reduced NADPH
					Or NADH
Aldehyde dehydrogenase (EC 1.)
k0n0-235	Y09876	Aldehyde dehydrogenase ( Nicotiana tabacum );	1.1e−07	152-191	Oxidation of aldehyde
passed		Length = 542			product of FMO to
					carboxylic acid
Permease
r0v0-239	S64084	Choline transport protein, yeast	9.3e−05	337-397	Transport of 2-OP1 into the
		Length = 563			cytoplasm
r0w0-424	S51169	amino acid transporter AAP4- Arabidopsis	0.98	8-76	Transport of 2-OP1 into the
w0h0-268		thaliana			cytoplasm
		len = 466
r0w0-205	P53744	KAPA/DAPA permease, yeast BIO5	2.1e−07	446-488	Transport of 2-OP1 into the
p0t0-308		Length = 561			cytoplasm
(contig)
Transmembrane pump (P-glycoprotein homolog)
r0g1-420	S20548	Leptomycin resistance protein, pmdl,	1.8 e−37	1255-1359	Transmembrane pump that
		Schizosaccharomyces pombe .		or	removes FB1 from the
		Length = 1362		564-668	cytoplasm as a means of
					protection against its toxic
					activity
g0s0-142		Leptomycin resistance protein, pmdl,		527-588	Transmembrane pump that
		Schizosaccharomyces pombe .			removes FB1 from the
		Length = 1362			cytoplasm as a means of
					protection against its toxic
					activity
10c0-129		Leptomycin resistance protein, pmdl,
		Schizosaccharomyces pombe .
		Length = 1362
r0s0-180		Leptomycin resistance protein, pmdl,		959-1009	Transmembrane pump that
		Schizosaccharomyces pombe .			removes FB1 from the
		Length = 1362			cytoplasm as a means of
					protection against its toxic
					activity
r0c0-193		Leptomycin resistance protein, pmdl,		885-945	Transmembrance pump that
		Schizosaccharomyces pombe .			removes FB1 from the
		Length = 1362			cytoplasm as a means of
					protection against its toxic
					activity
r0s0-330		Leptomycin resistance protein, pmdl,		1024-1110	Transmembrance pump that
		Schizosaccharomyces pombe .			removes FB1 from the
		Length = 1362			cytoplasm as a means of
					protection against its toxic
					activity
Loc0-129	S20548	Leptomycin resistance protein, pmdl,	.0082	949-988	Transmembrance pump that
		Schizosaccharomyces pombe .			removes FB1 from the
		Length = 1362			cytoplasm as a means of
					protection against its toxic
					activity
r0h1-262		Leptomycin resistance protein, pmdl,		1135-1218	Transmembrane pump that
		Schizosaccharomyces pombe .			removes FB1 from the
		Length = 1362			cytoplasm as a means of
					protection against its toxic
					activity
i0c0-116	e219956	ATP binding cassette multidrug transporter,		1026-1114	Transmembrane pump that
		Emericella nidulans Length = 1466			removes FB1 from the
					cytoplasm as a means of
					protection against its toxic
					activity

TABLE 3
Cloned
Bands	Homology, Comments	Predicted function	Predicted Product
1. Transmembrane pump (P-glycoprotein homolog)
r0g1-420 g0s0-142 10c0-129 r0s0-180 r0c0-193 r0s0-330 r0h1-262 i0c0-116	Homology to Leptomycin resistance protein, Pmd1, Schizosaccharomyces pombe , Length = 1362, or other ABC transporter gene family member. { } All 9 bands show homology to members of the ABC transporter superfamily.	FB1 Pump: Transmembrane pump that removes FB1 from the cytoplasm as a means of protection against its toxicity
2. Small Molecule Permease
r0v0-239 r0w0-205 p0t0-308.4 r0w0-424? w0h0-268?	Homology to choline transport protein, yeast Length = 563 { }Two bands (r0w0-205 and p0t0-308) contig with each other.	2-OP permease: Transport of 2- OP and/or AP1 into the cytoplasm
3. Flavin Monooxygenase (EC 1.14.13.22)
m1a0-388	Homology to cyclohexanone monooxygenase., Acinetobacter. Oxidation of ketone resulting in carbon-carbon bond breakage to form aldehyde. Utilizes NAD+ or NADP+	2-OP monooxygenase: Intracellular oxidation of 2-OP1 to a hydroxy aldehyde (HA-1) plus acetic acid
4. Aldehyde dehydrogenase
k0n0-235	Homology to aldehyde dehydrogenase ( Nicotiana tabacum ); Length = 542	HA-1 deydrogenase: Oxidation of aldehyde product of FMO to a hydroxy carboxylic acid (HCA-1)

Using sequence derived from each clone, a partial cDNA was obtained by 3′ and 5′ RACE-PCR (Chenchik et al. (1995) CLONTECHniques X 1:5-8); Chenchik et al. (1996) in A Laboratory Guide to RNA: Isolation, Analysis, and Synthesis, ed. Krieg (Wiley-Liss, Inc.), pp. 273-321. A RACE cloning kit from CLONTECH was used to obtain the RACE amplicons. Briefly, poly A+ RNA is transcribed to make first strand cDNA using a “lock-docking” poly T, cDNA synthesis primer, the second strand is synthesized, and the Marathon cDNA adaptor is ligated to both ends of the ds cDNA. Diluted template is then used with the Marathon adapter primer and in separate reactions either a 5′ Gene Specific Primer (GSP) or a 3′GSP is used to produce the 3′ or 5′ RACE amplicon. After characterization of the RACE product(s) and sequencing, full-length cDNAs may be generated by 1) end-to-end PCR using distal 5′ and 3′ GSPs with the adapter-ligated ds cDNA as template, or 2) the cloned 5′ and 3′-RACE fragments may be digested with a restriction enzyme that cuts uniquely in the region of overlap, and the fragments isolated and ligated. Subsequently, the RACE-generated full-length cDNAs from 1) and 2) may be cloned into a suitable vector.

EXAMPLE 6

Pichia Expression of Degradative Enzymes

For cloning into Pichia pastoris expression vector, pPicZalphaA, oligonucleotide primers were designed that contain a 22 bp overlap of the 5′ end (sense strand) and 3′ end (antisense strand), respectively of the open reading frame of the degradative nucleotide of interest, 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/NotI 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. After the generation of the 5′ and 3′ RACE products, the resulting band was cloned into EcoRI/NotI digested pPicZalphaA plasmid.

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 (negative control, no expression) or without an intron (capable of making an active protein). The Pichia culture fluids and pellets are assayed for enzyme activity as described earlier. The six day culture fluids from the same cultures are used to spike with crude fungal enzyme for positive controls.

The sample 50 μl cell pellets are resuspended in 150 μl cold 50 mM Na-phosphate, pH8.0 and divided into two fresh 500 μL 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:

1) pellet suspensions (“PELL”); 10 uL

2) lysate suspensions (“LYS”); 10 uL

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

4) positive control-used crude fungal enzyme undiluted; 10 uL

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

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 for 6 days

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 PhosphorImager (Molecular Dynamics) to develop images

EXAMPLE 7

Expression of Degradative Enzymes in E. coli

A vector for expressing the enzymes in E. coli 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 Iq 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 is cloned into the 5′ EcoRI site and a 3′ NotI site allowing in-frame expression of the fusion peptide. The generation of such an insert is described in the previous example.

E. coli is transformed with the vector containing the coding sequence for the degradative 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. coil is induced by addition of IPTG (isopropyl b-D-thiogalactopyranoside). Samples of soluble extract and Samples of insoluble inclusion bodies are tested for enzyme activity as described in Example 7.

EXAMPLE 8

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. 2 ). 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
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 3.5 g/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	4.000	g
(SIGMA C-1416)
Eriksson's Vitamin Mix	1.000	ml
(1000X SIGMA-1511
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	4.000	g
(SIGMA C-1416)
Eriksson's Vitamin Mix	1.000	ml
(1000X SIGMA-1511
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.

33 1 1691 DNA Exophiala spinifera misc_feature (0)...(0) flavin monooxygenase with intron 1atgtcggcca ccagcaactc cagaggcgat tgttccgtcg catgcgacgc catcatcgtt 60ggagccggcc tcagcggcat ctctgctgtg tacaaattgc gaaagctcag actcaacgcc 120aaaatcttcg agggagcccc cgattttggc ggcgtctggc actggaaccg ctaccctggc 180gctcgtgttg attcggagac gcccttctac caactgaaca ttcccgaagt atggaaagac 240tggacctggt cttgccgcta tcctgaccag aaagagttgc tgtcatatgt tcaccactgt 300gacaagatcc ggggcttgag aaaagacgtc tacttcggag ctgaggtggt tgatgcgcgg 360tatgccagag atctgggcac ctggactgtc aagacgtcgg ctggccatgt tgcgacggca 420aagtatctca ttctcgctac ggggttgctc cacaggaagc acactcccgc actccccggc 480ctcgccgatt tcaacgggaa ggtgattcat tcgagtgcct ggcacgaaga cttcgacgca 540gagggccaga gagtcgccgt catcggtgcc ggggccacaa gcatccagat tgttcaggag 600ttggccaaga aggctgacca ggtaaccatg tttatgcgaa ggccgagcta ttgtctgccc 660atgcggcaac gaacgatgga taggaacgaa cagacagcct ggaaggccta ctaccccacg 720ctgtttgaag cgagtcgaaa gtctcggatt ggattcccgg tccaggcacc gtcggttggc 780atctttgaag tcagccccga gcagcgggag gcctatttcg aagagttgtg ggagcgtggg 840gcctttaatt ttcttgcttg ccagtaccga gaagtcatgg ttgacaaaaa ggccaaccga 900ctggtctatg acttctgggc caaaaagact cgatctcgta tcgtcaatcc ggcaaagaga 960gatctcatgg ctcctctgga gccgccgtac tggttcggta ccaagcgctc cccactggag 1020agcgactact acgaaatgct ggacaagccg agcgtcgaaa ttgtgaatct agaacaatcg 1080cccattgtgg ctgttacaaa gacaggtgtg ctcttgagtg acggcagcaa gagggaatgc 1140gacacgatcg tgctggcgac gggtttcgac agtttcactg gctcgtgagt gtgctcgatc 1200atggctccga gtccggacgt ttggctgacc ttgaaagatt gacacatatg ggcttgaaaa 1260acaagcacgg agtggacctg aaggaggtgt ggaaagatgg catatctact tatatgggag 1320tcttctctca tggcttcccc aatgccttct tcgtcgccac ggctcaagcc ccgaccgtcc 1380tttccaacgg cccaacgatc atagaaaccc aagtcgactt gatcgccgat acaattgcaa 1440agttggaggc cgagcacgcc acgtccgttg aggcgacgaa atcagcacaa gaggcatggt 1500cgattatgat tgccaagatg aacgagcaca ctctgttccc cttgacggat tcgtggtgga 1560ctggaggcaa catccctggg aaagcaacac gtgctttaac cttcataggc gggattgctc 1620tctatgagca gatctgtcaa gagaaggtgg ccaattggga tgggtttgat gtgcttcatg 1680ctccctgcta a 1691 2 1638 DNA Exophiala spinifera CDS (1)...(1638) misc_feature (0)...(0) flavin monooxygenase, fully spliced 2atg tcg gcc acc agc aac tcc aga ggc gat tgt tcc gtc gca tgc gac 48Met Ser Ala Thr Ser Asn Ser Arg Gly Asp Cys Ser Val Ala Cys Asp 1 5 10 15gcc atc atc gtt gga gcc ggc ctc agc ggc atc tct gct gtg tac aaa 96Ala Ile Ile Val Gly Ala Gly Leu Ser Gly Ile Ser Ala Val Tyr Lys 20 25 30ttg cga aag ctc aga ctc aac gcc aaa atc ttc gag gga gcc ccc gat 144Leu Arg Lys Leu Arg Leu Asn Ala Lys Ile Phe Glu Gly Ala Pro Asp 35 40 45ttt ggc ggc gtc tgg cac tgg aac cgc tac cct ggc gct cgt gtt gat 192Phe Gly Gly Val Trp His Trp Asn Arg Tyr Pro Gly Ala Arg Val Asp 50 55 60tcg gag acg ccc ttc tac caa ctg aac att ccc gaa gta tgg aaa gac 240Ser Glu Thr Pro Phe Tyr Gln Leu Asn Ile Pro Glu Val Trp Lys Asp 65 70 75 80tgg acc tgg tct tgc cgc tat cct gac cag aaa gag ttg ctg tca tat 288Trp Thr Trp Ser Cys Arg Tyr Pro Asp Gln Lys Glu Leu Leu Ser Tyr 85 90 95gtt cac cac tgt gac aag atc cgg ggc ttg aga aaa gac gtc tac ttc 336Val His His Cys Asp Lys Ile Arg Gly Leu Arg Lys Asp Val Tyr Phe 100 105 110gga gct gag gtg gtt gat gcg cgg tat gcc aga gat ctg ggc acc tgg 384Gly Ala Glu Val Val Asp Ala Arg Tyr Ala Arg Asp Leu Gly Thr Trp 115 120 125act gtc aag acg tcg gct ggc cat gtt gcg acg gca aag tat ctc att 432Thr Val Lys Thr Ser Ala Gly His Val Ala Thr Ala Lys Tyr Leu Ile 130 135 140ctc gct acg ggg ttg ctc cac agg aag cac act ccc gca ctc ccc ggc 480Leu Ala Thr Gly Leu Leu His Arg Lys His Thr Pro Ala Leu Pro Gly145 150 155 160ctc gcc gat ttc aac ggg aag gtg att cat tcg agt gcc tgg cac gaa 528Leu Ala Asp Phe Asn Gly Lys Val Ile His Ser Ser Ala Trp His Glu 165 170 175gac ttc gac gca gag ggc cag aga gtc gcc gtc atc ggt gcc ggg gcc 576Asp Phe Asp Ala Glu Gly Gln Arg Val Ala Val Ile Gly Ala Gly Ala 180 185 190aca agc atc cag att gtt cag gag ttg gcc aag aag gct gac cag gta 624Thr Ser Ile Gln Ile Val Gln Glu Leu Ala Lys Lys Ala Asp Gln Val 195 200 205acc atg ttt atg cga agg ccg agc tat tgt ctg ccc atg cgg caa cga 672Thr Met Phe Met Arg Arg Pro Ser Tyr Cys Leu Pro Met Arg Gln Arg 210 215 220acg atg gat agg aac gaa cag aca gcc tgg aag gcc tac tac ccc acg 720Thr Met Asp Arg Asn Glu Gln Thr Ala Trp Lys Ala Tyr Tyr Pro Thr225 230 235 240ctg ttt gaa gcg agt cga aag tct cgg att gga ttc ccg gtc cag gca 768Leu Phe Glu Ala Ser Arg Lys Ser Arg Ile Gly Phe Pro Val Gln Ala 245 250 255ccg tcg gtt ggc atc ttt gaa gtc agc ccc gag cag cgg gag gcc tat 816Pro Ser Val Gly Ile Phe Glu Val Ser Pro Glu Gln Arg Glu Ala Tyr 260 265 270ttc gaa gag ttg tgg gag cgt ggg gcc ttt aat ttt ctt gct tgc cag 864Phe Glu Glu Leu Trp Glu Arg Gly Ala Phe Asn Phe Leu Ala Cys Gln 275 280 285tac cga gaa gtc atg gtt gac aaa aag gcc aac cga ctg gtc tat gac 912Tyr Arg Glu Val Met Val Asp Lys Lys Ala Asn Arg Leu Val Tyr Asp 290 295 300ttc tgg gcc aaa aag act cga tct cgt atc gtc aat ccg gca aag aga 960Phe Trp Ala Lys Lys Thr Arg Ser Arg Ile Val Asn Pro Ala Lys Arg305 310 315 320gat ctc atg gct cct ctg gag ccg ccg tac tgg ttc ggt acc aag cgc 1008Asp Leu Met Ala Pro Leu Glu Pro Pro Tyr Trp Phe Gly Thr Lys Arg 325 330 335tcc cca ctg gag agc gac tac tac gaa atg ctg gac aag ccg agc gtc 1056Ser Pro Leu Glu Ser Asp Tyr Tyr Glu Met Leu Asp Lys Pro Ser Val 340 345 350gaa att gtg aat cta gaa caa tcg ccc att gtg gct gtt aca aag aca 1104Glu Ile Val Asn Leu Glu Gln Ser Pro Ile Val Ala Val Thr Lys Thr 355 360 365ggt gtg ctc ttg agt gac ggc agc aag agg gaa tgc gac acg atc gtg 1152Gly Val Leu Leu Ser Asp Gly Ser Lys Arg Glu Cys Asp Thr Ile Val 370 375 380ctg gcg acg ggt ttc gac agt ttc act ggc tca ttg aca cat atg ggc 1200Leu Ala Thr Gly Phe Asp Ser Phe Thr Gly Ser Leu Thr His Met Gly385 390 395 400ttg aaa aac aag cac gga gtg gac ctg aag gag gtg tgg aaa gat ggc 1248Leu Lys Asn Lys His Gly Val Asp Leu Lys Glu Val Trp Lys Asp Gly 405 410 415ata tct act tat atg gga gtc ttc tct cat ggc ttc ccc aat gcc ttc 1296Ile Ser Thr Tyr Met Gly Val Phe Ser His Gly Phe Pro Asn Ala Phe 420 425 430ttc gtc gcc acg gct caa gcc ccg acc gtc ctt tcc aac ggc cca acg 1344Phe Val Ala Thr Ala Gln Ala Pro Thr Val Leu Ser Asn Gly Pro Thr 435 440 445atc ata gaa acc caa gtc gac ttg atc gcc gat aca att gca aag ttg 1392Ile Ile Glu Thr Gln Val Asp Leu Ile Ala Asp Thr Ile Ala Lys Leu 450 455 460gag gcc gag cac gcc acg tcc gtt gag gcg acg aaa tca gca caa gag 1440Glu Ala Glu His Ala Thr Ser Val Glu Ala Thr Lys Ser Ala Gln Glu465 470 475 480gca tgg tcg att atg att gcc aag atg aac gag cac act ctg ttc ccc 1488Ala Trp Ser Ile Met Ile Ala Lys Met Asn Glu His Thr Leu Phe Pro 485 490 495ttg acg gat tcg tgg tgg act gga ggc aac atc cct ggg aaa gca aca 1536Leu Thr Asp Ser Trp Trp Thr Gly Gly Asn Ile Pro Gly Lys Ala Thr 500 505 510cgt gct tta acc ttc ata ggc ggg att gct ctc tat gag cag atc tgt 1584Arg Ala Leu Thr Phe Ile Gly Gly Ile Ala Leu Tyr Glu Gln Ile Cys 515 520 525caa gag aag gtg gcc aat tgg gat ggg ttt gat gtg ctt cat gct ccc 1632Gln Glu Lys Val Ala Asn Trp Asp Gly Phe Asp Val Leu His Ala Pro 530 535 540tgc taa 1638Cys *545 3 545 PRT Exophiala spinifera 3Met Ser Ala Thr Ser Asn Ser Arg Gly Asp Cys Ser Val Ala Cys Asp 1 5 10 15Ala Ile Ile Val Gly Ala Gly Leu Ser Gly Ile Ser Ala Val Tyr Lys 20 25 30Leu Arg Lys Leu Arg Leu Asn Ala Lys Ile Phe Glu Gly Ala Pro Asp 35 40 45Phe Gly Gly Val Trp His Trp Asn Arg Tyr Pro Gly Ala Arg Val Asp 50 55 60Ser Glu Thr Pro Phe Tyr Gln Leu Asn Ile Pro Glu Val Trp Lys Asp65 70 75 80Trp Thr Trp Ser Cys Arg Tyr Pro Asp Gln Lys Glu Leu Leu Ser Tyr 85 90 95Val His His Cys Asp Lys Ile Arg Gly Leu Arg Lys Asp Val Tyr Phe 100 105 110Gly Ala Glu Val Val Asp Ala Arg Tyr Ala Arg Asp Leu Gly Thr Trp 115 120 125Thr Val Lys Thr Ser Ala Gly His Val Ala Thr Ala Lys Tyr Leu Ile 130 135 140Leu Ala Thr Gly Leu Leu His Arg Lys His Thr Pro Ala Leu Pro Gly145 150 155 160Leu Ala Asp Phe Asn Gly Lys Val Ile His Ser Ser Ala Trp His Glu 165 170 175Asp Phe Asp Ala Glu Gly Gln Arg Val Ala Val Ile Gly Ala Gly Ala 180 185 190Thr Ser Ile Gln Ile Val Gln Glu Leu Ala Lys Lys Ala Asp Gln Val 195 200 205Thr Met Phe Met Arg Arg Pro Ser Tyr Cys Leu Pro Met Arg Gln Arg 210 215 220Thr Met Asp Arg Asn Glu Gln Thr Ala Trp Lys Ala Tyr Tyr Pro Thr225 230 235 240Leu Phe Glu Ala Ser Arg Lys Ser Arg Ile Gly Phe Pro Val Gln Ala 245 250 255Pro Ser Val Gly Ile Phe Glu Val Ser Pro Glu Gln Arg Glu Ala Tyr 260 265 270Phe Glu Glu Leu Trp Glu Arg Gly Ala Phe Asn Phe Leu Ala Cys Gln 275 280 285Tyr Arg Glu Val Met Val Asp Lys Lys Ala Asn Arg Leu Val Tyr Asp 290 295 300Phe Trp Ala Lys Lys Thr Arg Ser Arg Ile Val Asn Pro Ala Lys Arg305 310 315 320Asp Leu Met Ala Pro Leu Glu Pro Pro Tyr Trp Phe Gly Thr Lys Arg 325 330 335Ser Pro Leu Glu Ser Asp Tyr Tyr Glu Met Leu Asp Lys Pro Ser Val 340 345 350Glu Ile Val Asn Leu Glu Gln Ser Pro Ile Val Ala Val Thr Lys Thr 355 360 365Gly Val Leu Leu Ser Asp Gly Ser Lys Arg Glu Cys Asp Thr Ile Val 370 375 380Leu Ala Thr Gly Phe Asp Ser Phe Thr Gly Ser Leu Thr His Met Gly385 390 395 400Leu Lys Asn Lys His Gly Val Asp Leu Lys Glu Val Trp Lys Asp Gly 405 410 415Ile Ser Thr Tyr Met Gly Val Phe Ser His Gly Phe Pro Asn Ala Phe 420 425 430Phe Val Ala Thr Ala Gln Ala Pro Thr Val Leu Ser Asn Gly Pro Thr 435 440 445Ile Ile Glu Thr Gln Val Asp Leu Ile Ala Asp Thr Ile Ala Lys Leu 450 455 460Glu Ala Glu His Ala Thr Ser Val Glu Ala Thr Lys Ser Ala Gln Glu465 470 475 480Ala Trp Ser Ile Met Ile Ala Lys Met Asn Glu His Thr Leu Phe Pro 485 490 495Leu Thr Asp Ser Trp Trp Thr Gly Gly Asn Ile Pro Gly Lys Ala Thr 500 505 510Arg Ala Leu Thr Phe Ile Gly Gly Ile Ala Leu Tyr Glu Gln Ile Cys 515 520 525Gln Glu Lys Val Ala Asn Trp Asp Gly Phe Asp Val Leu His Ala Pro 530 535 540Cys545 4 1464 DNA Exophiala spinifera CDS (1)...(1464) misc_feature (0)...(0) aldehyde dehydrogenase, fully spliced DNA 4atg gtt ctt tcg cct gac gaa tac aag agt gaa ctc ttc atc aac aat 48Met Val Leu Ser Pro Asp Glu Tyr Lys Ser Glu Leu Phe Ile Asn Asn 1 5 10 15gaa ttc gtc tcc tcc aag ggg tcc gag aga tta acg ctc acg aac ccg 96Glu Phe Val Ser Ser Lys Gly Ser Glu Arg Leu Thr Leu Thr Asn Pro 20 25 30tgg gac gaa tcc acc gtt gcc act gat gtt cac gtg gcc aac gcg gcc 144Trp Asp Glu Ser Thr Val Ala Thr Asp Val His Val Ala Asn Ala Ala 35 40 45gat gtc gac agt gca gta gcc gct tcg gtg cag gcg gtc aaa aag ggc 192Asp Val Asp Ser Ala Val Ala Ala Ser Val Gln Ala Val Lys Lys Gly 50 55 60cca tgg aag aag ttc aca ggt gca caa cgc gcg gcg tgc atg ctt aag 240Pro Trp Lys Lys Phe Thr Gly Ala Gln Arg Ala Ala Cys Met Leu Lys 65 70 75 80ttc gcg gac ctc gcc gag aag aac gcc gag aag ctc gct cgt ctg gag 288Phe Ala Asp Leu Ala Glu Lys Asn Ala Glu Lys Leu Ala Arg Leu Glu 85 90 95tcg ctg ccc acc ggt aga ccg gtg tcg atg atc act cat ttc gac att 336Ser Leu Pro Thr Gly Arg Pro Val Ser Met Ile Thr His Phe Asp Ile 100 105 110cca aac atg gtc tcc gtg ttt cgc tac tat gca ggc tgg gcc gac aag 384Pro Asn Met Val Ser Val Phe Arg Tyr Tyr Ala Gly Trp Ala Asp Lys 115 120 125atc gcc gga aag acc ttt ccc gag gac aac ggc aag ccg aat tgg cgt 432Ile Ala Gly Lys Thr Phe Pro Glu Asp Asn Gly Lys Pro Asn Trp Arg 130 135 140tac gag ccg atg ggg gtg tgt gct ggt att gcc agc tgg aac gcg act 480Tyr Glu Pro Met Gly Val Cys Ala Gly Ile Ala Ser Trp Asn Ala Thr145 150 155 160ttt ctt tac gtc ggc tgg aag ata gcc ccc gcc ctc gcc gcc ggc tgc 528Phe Leu Tyr Val Gly Trp Lys Ile Ala Pro Ala Leu Ala Ala Gly Cys 165 170 175tcc ttc atc ttc aaa gcc tcg gag aaa tcc ccg ctg ggc gtt ctg ggc 576Ser Phe Ile Phe Lys Ala Ser Glu Lys Ser Pro Leu Gly Val Leu Gly 180 185 190ctc gct cct ctc ttc gca gaa gcc gga ttc cct cct gga gtc gtg cag 624Leu Ala Pro Leu Phe Ala Glu Ala Gly Phe Pro Pro Gly Val Val Gln 195 200 205ttc ctc act gga gca cga gtg acg ggt gaa gca ttg gcg tcg cac atg 672Phe Leu Thr Gly Ala Arg Val Thr Gly Glu Ala Leu Ala Ser His Met 210 215 220gac att gcg aag atc agc ttc aca aga tct gtc ggc ggt ggc cgc gcc 720Asp Ile Ala Lys Ile Ser Phe Thr Arg Ser Val Gly Gly Gly Arg Ala225 230 235 240gtc aag caa gca aca ctc aag tcc aac atg aag cgc gtc act cta gaa 768Val Lys Gln Ala Thr Leu Lys Ser Asn Met Lys Arg Val Thr Leu Glu 245 250 255ctg ggg gaa aag cca acc atc gtc ttc aac gaa gct cct ctc gaa cgg 816Leu Gly Glu Lys Pro Thr Ile Val Phe Asn Glu Ala Pro Leu Glu Arg 260 265 270cag tcg ggg gaa tcg gca aag gat ttc tca aaa ttc ggg caa att tgg 864Gln Ser Gly Glu Ser Ala Lys Asp Phe Ser Lys Phe Gly Gln Ile Trp 275 280 285gtc ccc ccc tcc tgt ttg cta gtg caa tgg gga aat tta gcg gag aaa 912Val Pro Pro Ser Cys Leu Leu Val Gln Trp Gly Asn Leu Ala Glu Lys 290 295 300ttc cat gga gtc cgt cat ggc tca ttt gga ggc tgt cag aga tgg ctt 960Phe His Gly Val Arg His Gly Ser Phe Gly Gly Cys Gln Arg Trp Leu305 310 315 320ggc cag aac cca ttg gaa ccc aag agg acg cat ggt ccc ttc gtc gac 1008Gly Gln Asn Pro Leu Glu Pro Lys Arg Thr His Gly Pro Phe Val Asp 325 330 335aag tcc cag tac gac aga gtc ttg ggt aac att gac gtt ggc aag gat 1056Lys Ser Gln Tyr Asp Arg Val Leu Gly Asn Ile Asp Val Gly Lys Asp 340 345 350acc gcg cag ctc ctc act ggc gtt ggt aga aag ggc gac aag gga ttc 1104Thr Ala Gln Leu Leu Thr Gly Val Gly Arg Lys Gly Asp Lys Gly Phe 355 360 365gcg att gaa ccg acg ata ttt gtc aat ccc aaa cca ggc agc aaa att 1152Ala Ile Glu Pro Thr Ile Phe Val Asn Pro Lys Pro Gly Ser Lys Ile 370 375 380tgg ttt gag gag atc ttt ggc ccc gtc ttg tcc att aag acg ttc aag 1200Trp Phe Glu Glu Ile Phe Gly Pro Val Leu Ser Ile Lys Thr Phe Lys385 390 395 400acg gaa gaa gag gcc att gag att gcc aat gac acg act tat ggg cta 1248Thr Glu Glu Glu Ala Ile Glu Ile Ala Asn Asp Thr Thr Tyr Gly Leu 405 410 415gcc tcg gtc att tat acc aaa tct ctc aac agg ggt ctc cgt gtc tcg 1296Ala Ser Val Ile Tyr Thr Lys Ser Leu Asn Arg Gly Leu Arg Val Ser 420 425 430tcg gcg ctc gag acc ggt ggc gtc tcg atc aac ttc ccc ttt atc ccc 1344Ser Ala Leu Glu Thr Gly Gly Val Ser Ile Asn Phe Pro Phe Ile Pro 435 440 445gag aca caa act ccg ttt ggc ggc atg aaa caa tcg ggc tca ggc aga 1392Glu Thr Gln Thr Pro Phe Gly Gly Met Lys Gln Ser Gly Ser Gly Arg 450 455 460gag cta ggc gaa gaa ggg ctc aag gcg tac ttg gag ccc aag acc att 1440Glu Leu Gly Glu Glu Gly Leu Lys Ala Tyr Leu Glu Pro Lys Thr Ile465 470 475 480aat atc cac gtc aac ata gag tga 1464Asn Ile His Val Asn Ile Glu * 485 5 487 PRT Exophiala spinifera 5Met Val Leu Ser Pro Asp Glu Tyr Lys Ser Glu Leu Phe Ile Asn Asn 1 5 10 15Glu Phe Val Ser Ser Lys Gly Ser Glu Arg Leu Thr Leu Thr Asn Pro 20 25 30Trp Asp Glu Ser Thr Val Ala Thr Asp Val His Val Ala Asn Ala Ala 35 40 45Asp Val Asp Ser Ala Val Ala Ala Ser Val Gln Ala Val Lys Lys Gly 50 55 60Pro Trp Lys Lys Phe Thr Gly Ala Gln Arg Ala Ala Cys Met Leu Lys65 70 75 80Phe Ala Asp Leu Ala Glu Lys Asn Ala Glu Lys Leu Ala Arg Leu Glu 85 90 95Ser Leu Pro Thr Gly Arg Pro Val Ser Met Ile Thr His Phe Asp Ile 100 105 110Pro Asn Met Val Ser Val Phe Arg Tyr Tyr Ala Gly Trp Ala Asp Lys 115 120 125Ile Ala Gly Lys Thr Phe Pro Glu Asp Asn Gly Lys Pro Asn Trp Arg 130 135 140Tyr Glu Pro Met Gly Val Cys Ala Gly Ile Ala Ser Trp Asn Ala Thr145 150 155 160Phe Leu Tyr Val Gly Trp Lys Ile Ala Pro Ala Leu Ala Ala Gly Cys 165 170 175Ser Phe Ile Phe Lys Ala Ser Glu Lys Ser Pro Leu Gly Val Leu Gly 180 185 190Leu Ala Pro Leu Phe Ala Glu Ala Gly Phe Pro Pro Gly Val Val Gln 195 200 205Phe Leu Thr Gly Ala Arg Val Thr Gly Glu Ala Leu Ala Ser His Met 210 215 220Asp Ile Ala Lys Ile Ser Phe Thr Arg Ser Val Gly Gly Gly Arg Ala225 230 235 240Val Lys Gln Ala Thr Leu Lys Ser Asn Met Lys Arg Val Thr Leu Glu 245 250 255Leu Gly Glu Lys Pro Thr Ile Val Phe Asn Glu Ala Pro Leu Glu Arg 260 265 270Gln Ser Gly Glu Ser Ala Lys Asp Phe Ser Lys Phe Gly Gln Ile Trp 275 280 285Val Pro Pro Ser Cys Leu Leu Val Gln Trp Gly Asn Leu Ala Glu Lys 290 295 300Phe His Gly Val Arg His Gly Ser Phe Gly Gly Cys Gln Arg Trp Leu305 310 315 320Gly Gln Asn Pro Leu Glu Pro Lys Arg Thr His Gly Pro Phe Val Asp 325 330 335Lys Ser Gln Tyr Asp Arg Val Leu Gly Asn Ile Asp Val Gly Lys Asp 340 345 350Thr Ala Gln Leu Leu Thr Gly Val Gly Arg Lys Gly Asp Lys Gly Phe 355 360 365Ala Ile Glu Pro Thr Ile Phe Val Asn Pro Lys Pro Gly Ser Lys Ile 370 375 380Trp Phe Glu Glu Ile Phe Gly Pro Val Leu Ser Ile Lys Thr Phe Lys385 390 395 400Thr Glu Glu Glu Ala Ile Glu Ile Ala Asn Asp Thr Thr Tyr Gly Leu 405 410 415Ala Ser Val Ile Tyr Thr Lys Ser Leu Asn Arg Gly Leu Arg Val Ser 420 425 430Ser Ala Leu Glu Thr Gly Gly Val Ser Ile Asn Phe Pro Phe Ile Pro 435 440 445Glu Thr Gln Thr Pro Phe Gly Gly Met Lys Gln Ser Gly Ser Gly Arg 450 455 460Glu Leu Gly Glu Glu Gly Leu Lys Ala Tyr Leu Glu Pro Lys Thr Ile465 470 475 480Asn Ile His Val Asn Ile Glu 485 6 1764 DNA Exophiala spinifera misc_feature (0)...(0) permease, partially spliced cDNA 6aactatggac tccagaccaa gtggatacgg cgagaaaggc gggacaaggc agacaacgaa 60gaacacagag acggcggcgg caggtggtgc gtccgagtcc ctgaacgttc ctctggagaa 120gaaacaattt ggcaccatca ccatcgtgtc cttggccttt gtgatttgca acagttgggc 180tggtatctca ggcagtctcc agctcgccct actagcgggg gggcccgtca ctctccttta 240cggcatccta atcagtactc tcgtctacat ctgcatcgct ttctcattag ccgaactgac 300cagcgtctac ccgactgccg gtggccaata tcattttgcg tcgatcctgg caccaaaatc 360aatcaatcgg agcatttcat acgtgtgcgg actcgtgtcg ttgctttcat ggatcgctat 420cggaagctca gtgaccatga tacctgctca acagatcccg gcgctgatag ccgcctatag 480tcacacatac tcccaggatt cgtggcatgt cttcctcatc tacgagggag tcgcgctggt 540ggtgctcttg ttcaacttgt ttgccctgaa aagaaaccct tgggttcatg aaatcggatt 600cggcctcacg atcgctctct tcgtgatctc ctttatcgcc attctagcgc ggtccaaccc 660caaggctcca aactcacagg tatggactgc ttggagcaac tatactggct ggtccgacgg 720cgtctgcttc atcctgggcc tttcgacatc ctgcttcatg ttcattggct tggacgcagc 780aatgcatctg gctgaagaat gcacagatgc tgctcgtacg gtacccaaag cagtggtcag 840tgcaatcata attggcttct gcaccgcctt tccatataca atcgcagttc tgtatggaat 900tacagatctc gactctattc taagttccgc cggctatatt ccattcgaga caatgacgca 960gtcccttcgg tcgctcagtt ttgcaacggt cctctcatgt ggcggtatcg tgatggcctt 1020cttcgccctc aacgctgtac aagagactgc gtctcgactc acctggagct ttgcccggga 1080caatgggctg gtattttcca ctcatctcga acgcattcat ccccgctggc aagttcctgt 1140ttggtctcta ttcgcgacct ggggaattct ggccacatgc ggatgtatat ttctaggttc 1200tagcacagct ttcaatgcct tggtcaattc cgccgttgta ctccagcaac tctccttcct 1260gatcccaatc gccctactcc tctaccaaaa gcgagatcca aagttcttgc cgagcactcg 1320tgcttttgtg ttaccgcgtg gaatcgggtt tctggtcaat gtgctagcgg tggtcttcac 1380gtccgtcacc actgtgtttt tcagcttccc actgaccgtg cctacggccg cgtcaaccat 1440gaattacaca agtgcgatta taggcgttgc acttgctctt ggtgtcttga actgggtcgt 1500gcatgccagg aagcattatc agggacccca cttggagctt gacggacggg tcgtcggagc 1560agaatttcaa gttgggccat gaattggacg aaatggagac gcgtgtgcaa tgtcaaaaat 1620tgctggggtg gtactgagag tctggattag ctgcaacgcg ggacaaccga gggtagaaca 1680ctctgcaatc gagcaggaca atatcaatta ggcaachasv caaaaaaaaa aaaaaaaaaa 1740aaaaaagcgg ccgctgaatt ctag 1764 7 1578 DNA Exophiala spinifera CDS (1)...(1578) misc_feature (0)...(0) permease, fully spliced cDNA 7atg gac tcc aga cca agt gga tac ggc gag aaa ggc ggg aca agg cag 48Met Asp Ser Arg Pro Ser Gly Tyr Gly Glu Lys Gly Gly Thr Arg Gln 1 5 10 15aca acg aag aac aca gag acg gcg gcg gca ggt ggt gcg tcc gag tcc 96Thr Thr Lys Asn Thr Glu Thr Ala Ala Ala Gly Gly Ala Ser Glu Ser 20 25 30ctg aac gtt cct ctg gag aag aaa caa ttt ggc acc atc acc atc gtg 144Leu Asn Val Pro Leu Glu Lys Lys Gln Phe Gly Thr Ile Thr Ile Val 35 40 45tcc ttg gcc ttt gtg att tgc aac agt tgg gct ggt atc tca ggc agt 192Ser Leu Ala Phe Val Ile Cys Asn Ser Trp Ala Gly Ile Ser Gly Ser 50 55 60ctc cag ctc gcc cta cta gcg ggg ggg ccc gtc act ctc ctt tac ggc 240Leu Gln Leu Ala Leu Leu Ala Gly Gly Pro Val Thr Leu Leu Tyr Gly 65 70 75 80atc cta atc agt act ctc gtc tac atc tgc atc gct ttc tca tta gcc 288Ile Leu Ile Ser Thr Leu Val Tyr Ile Cys Ile Ala Phe Ser Leu Ala 85 90 95gaa ctg acc agc gtc tac ccg act gcc ggt ggc caa tat cat ttt gcg 336Glu Leu Thr Ser Val Tyr Pro Thr Ala Gly Gly Gln Tyr His Phe Ala 100 105 110tcg atc ctg gca cca aaa tca atc aat cgg agc att tca tac gtg tgc 384Ser Ile Leu Ala Pro Lys Ser Ile Asn Arg Ser Ile Ser Tyr Val Cys 115 120 125gga ctc gtg tcg ttg ctt tca tgg atc gct atc gga agc tca gtg acc 432Gly Leu Val Ser Leu Leu Ser Trp Ile Ala Ile Gly Ser Ser Val Thr 130 135 140atg ata cct gct caa cag atc ccg gcg ctg ata gcc gcc tat agt cac 480Met Ile Pro Ala Gln Gln Ile Pro Ala Leu Ile Ala Ala Tyr Ser His145 150 155 160aca tac tcc cag gat tcg tgg cat gtc ttc ctc atc tac gag gga gtc 528Thr Tyr Ser Gln Asp Ser Trp His Val Phe Leu Ile Tyr Glu Gly Val 165 170 175gcg ctg gtg gtg ctc ttg ttc aac ttg ttt gcc ctg aaa aga aac cct 576Ala Leu Val Val Leu Leu Phe Asn Leu Phe Ala Leu Lys Arg Asn Pro 180 185 190tgg gtt cat gaa atc gga ttc ggc ctc acg atc gct ctc ttc gtg atc 624Trp Val His Glu Ile Gly Phe Gly Leu Thr Ile Ala Leu Phe Val Ile 195 200 205tcc ttt atc gcc att cta gcg cgg tcc aac ccc aag gct cca aac tca 672Ser Phe Ile Ala Ile Leu Ala Arg Ser Asn Pro Lys Ala Pro Asn Ser 210 215 220cag gta tgg act gct tgg agc aac tat act ggc tgg tcc gac ggc gtc 720Gln Val Trp Thr Ala Trp Ser Asn Tyr Thr Gly Trp Ser Asp Gly Val225 230 235 240tgc ttc atc ctg ggc ctt tcg aca tcc tgc ttc atg ttc att ggc ttg 768Cys Phe Ile Leu Gly Leu Ser Thr Ser Cys Phe Met Phe Ile Gly Leu 245 250 255gac gca gca atg cat ctg gct gaa gaa tgc aca gat gct gct cgt acg 816Asp Ala Ala Met His Leu Ala Glu Glu Cys Thr Asp Ala Ala Arg Thr 260 265 270gta ccc aaa gca gtg gtc agt gca atc ata att ggc ttc tgc acc gcc 864Val Pro Lys Ala Val Val Ser Ala Ile Ile Ile Gly Phe Cys Thr Ala 275 280 285ttt cca tat aca atc gca gtt ctg tat gga att aca gat ctc gac tct 912Phe Pro Tyr Thr Ile Ala Val Leu Tyr Gly Ile Thr Asp Leu Asp Ser 290 295 300att cta agt tcc gcc ggc tat att cca ttc gag aca atg acg cag tcc 960Ile Leu Ser Ser Ala Gly Tyr Ile Pro Phe Glu Thr Met Thr Gln Ser305 310 315 320ctt cgg tcg ctc agt ttt gca acg gtc ctc tca tgt ggc ggt atc gtg 1008Leu Arg Ser Leu Ser Phe Ala Thr Val Leu Ser Cys Gly Gly Ile Val 325 330 335atg gcc ttc ttc gcc ctc aac gct gta caa gag act gcg tct cga ctc 1056Met Ala Phe Phe Ala Leu Asn Ala Val Gln Glu Thr Ala Ser Arg Leu 340 345 350acc tgg agc ttt gcc cgg gac aat ggg ctg gta ttt tcc act cat ctc 1104Thr Trp Ser Phe Ala Arg Asp Asn Gly Leu Val Phe Ser Thr His Leu 355 360 365gaa cgc att cat ccc cgc tgg caa gtt cct gtt tgg tct cta ttc gcg 1152Glu Arg Ile His Pro Arg Trp Gln Val Pro Val Trp Ser Leu Phe Ala 370 375 380acc tgg gga att ctg gcc aca tgc gga tgt ata ttt cta ggt tct agc 1200Thr Trp Gly Ile Leu Ala Thr Cys Gly Cys Ile Phe Leu Gly Ser Ser385 390 395 400aca gct ttc aat gcc ttg gtc aat tcc gcc gtt gta ctc cag caa ctc 1248Thr Ala Phe Asn Ala Leu Val Asn Ser Ala Val Val Leu Gln Gln Leu 405 410 415tcc ttc ctg atc cca atc gcc cta ctc ctc tac caa aag cga gat cca 1296Ser Phe Leu Ile Pro Ile Ala Leu Leu Leu Tyr Gln Lys Arg Asp Pro 420 425 430aag ttc ttg ccg agc act cgt gct ttt gtg tta ccg cgt gga atc ggg 1344Lys Phe Leu Pro Ser Thr Arg Ala Phe Val Leu Pro Arg Gly Ile Gly 435 440 445ttt ctg gtc aat gtg cta gcg gtg gtc ttc acg tcc gtc acc act gtg 1392Phe Leu Val Asn Val Leu Ala Val Val Phe Thr Ser Val Thr Thr Val 450 455 460ttt ttc agc ttc cca ctg acc gtg cct acg gcc gcg tca acc atg aat 1440Phe Phe Ser Phe Pro Leu Thr Val Pro Thr Ala Ala Ser Thr Met Asn465 470 475 480tac aca agt gcg att ata ggc gtt gca ctt gct ctt ggt gtc ttg aac 1488Tyr Thr Ser Ala Ile Ile Gly Val Ala Leu Ala Leu Gly Val Leu Asn 485 490 495tgg gtc gtg cat gcc agg aag cat tat cag gga ccc cac ttg gag ctt 1536Trp Val Val His Ala Arg Lys His Tyr Gln Gly Pro His Leu Glu Leu 500 505 510gac gga cgg gtc gtc gga gca gaa ttt caa gtt ggg cca tga 1578Asp Gly Arg Val Val Gly Ala Glu Phe Gln Val Gly Pro * 515 520 525 8 525 PRT Exophiala spinifera 8Met Asp Ser Arg Pro Ser Gly Tyr Gly Glu Lys Gly Gly Thr Arg Gln 1 5 10 15Thr Thr Lys Asn Thr Glu Thr Ala Ala Ala Gly Gly Ala Ser Glu Ser 20 25 30Leu Asn Val Pro Leu Glu Lys Lys Gln Phe Gly Thr Ile Thr Ile Val 35 40 45Ser Leu Ala Phe Val Ile Cys Asn Ser Trp Ala Gly Ile Ser Gly Ser 50 55 60Leu Gln Leu Ala Leu Leu Ala Gly Gly Pro Val Thr Leu Leu Tyr Gly65 70 75 80Ile Leu Ile Ser Thr Leu Val Tyr Ile Cys Ile Ala Phe Ser Leu Ala 85 90 95Glu Leu Thr Ser Val Tyr Pro Thr Ala Gly Gly Gln Tyr His Phe Ala 100 105 110Ser Ile Leu Ala Pro Lys Ser Ile Asn Arg Ser Ile Ser Tyr Val Cys 115 120 125Gly Leu Val Ser Leu Leu Ser Trp Ile Ala Ile Gly Ser Ser Val Thr 130 135 140Met Ile Pro Ala Gln Gln Ile Pro Ala Leu Ile Ala Ala Tyr Ser His145 150 155 160Thr Tyr Ser Gln Asp Ser Trp His Val Phe Leu Ile Tyr Glu Gly Val 165 170 175Ala Leu Val Val Leu Leu Phe Asn Leu Phe Ala Leu Lys Arg Asn Pro 180 185 190Trp Val His Glu Ile Gly Phe Gly Leu Thr Ile Ala Leu Phe Val Ile 195 200 205Ser Phe Ile Ala Ile Leu Ala Arg Ser Asn Pro Lys Ala Pro Asn Ser 210 215 220Gln Val Trp Thr Ala Trp Ser Asn Tyr Thr Gly Trp Ser Asp Gly Val225 230 235 240Cys Phe Ile Leu Gly Leu Ser Thr Ser Cys Phe Met Phe Ile Gly Leu 245 250 255Asp Ala Ala Met His Leu Ala Glu Glu Cys Thr Asp Ala Ala Arg Thr 260 265 270Val Pro Lys Ala Val Val Ser Ala Ile Ile Ile Gly Phe Cys Thr Ala 275 280 285Phe Pro Tyr Thr Ile Ala Val Leu Tyr Gly Ile Thr Asp Leu Asp Ser 290 295 300Ile Leu Ser Ser Ala Gly Tyr Ile Pro Phe Glu Thr Met Thr Gln Ser305 310 315 320Leu Arg Ser Leu Ser Phe Ala Thr Val Leu Ser Cys Gly Gly Ile Val 325 330 335Met Ala Phe Phe Ala Leu Asn Ala Val Gln Glu Thr Ala Ser Arg Leu 340 345 350Thr Trp Ser Phe Ala Arg Asp Asn Gly Leu Val Phe Ser Thr His Leu 355 360 365Glu Arg Ile His Pro Arg Trp Gln Val Pro Val Trp Ser Leu Phe Ala 370 375 380Thr Trp Gly Ile Leu Ala Thr Cys Gly Cys Ile Phe Leu Gly Ser Ser385 390 395 400Thr Ala Phe Asn Ala Leu Val Asn Ser Ala Val Val Leu Gln Gln Leu 405 410 415Ser Phe Leu Ile Pro Ile Ala Leu Leu Leu Tyr Gln Lys Arg Asp Pro 420 425 430Lys Phe Leu Pro Ser Thr Arg Ala Phe Val Leu Pro Arg Gly Ile Gly 435 440 445Phe Leu Val Asn Val Leu Ala Val Val Phe Thr Ser Val Thr Thr Val 450 455 460Phe Phe Ser Phe Pro Leu Thr Val Pro Thr Ala Ala Ser Thr Met Asn465 470 475 480Tyr Thr Ser Ala Ile Ile Gly Val Ala Leu Ala Leu Gly Val Leu Asn 485 490 495Trp Val Val His Ala Arg Lys His Tyr Gln Gly Pro His Leu Glu Leu 500 505 510Asp Gly Arg Val Val Gly Ala Glu Phe Gln Val Gly Pro 515 520 525 9 3999 DNA Exophiala spinifera misc_feature (0)...(0) p-glycoprotein, with introns 9tatttsccat ctmckatgaa tggcagatga atcggagaaa cctcgaccaa accaagatgg 60cagtgagtcg tcctcacacc ctcccccaga aaaggaaacc gaaggcagta tttcagacta 120tctacgaatc ttcagatatg ccgacaaata cgactggact ctcaatgtca tcgcgctcat 180ctgcgccatc ggatccgggg cttcccttcc tctgatgtcg atcatcttcg gtagcttcac 240caacaagttc aacaattaca attcgggcga cgggagtcct gaagcgttca aggccgatgt 300ggatcatttc gtcctgtggt tcgtctacct ctttattggg aagtttgtcc tcacgtacgt 360ttccacggct gccattacca tttcagctat acgaaccact cgaactcttc gacgagtgtt 420ccttgaatgc accttgcggc aagaggtctg gcatttcgac aagcagagca atggagcaat 480cgccactcag gtcactacca atggcaaccg tatacaaaca ggtattgccg agaaattggt 540ctttaccgtg caggcacttt caatgttctt ttctgcattt gtggtcgctt tggcgtctca 600gtggaagcta gctttaatca ccatgtccgt catccctgcc attttcctgg tcaccggcat 660ctgcatagca attgatgccg ctcaggaggc caggatcacc aggatctact cacgcgccgc 720tgtcctcgca gaagaagtct tatcatccat ccggacagtc catgctttct acgcccagaa 780gaaaatggtc gaaaaatatg atgtcttttt gcagcaagca caccaagaag ggaagaagaa 840atcgccaaat tatggggtct tgttctcaac tgagtacttt tgcatttacg ctgctatcgc 900actgggcctt ttgggaaagg tttttcgcat gtatcagaat ggcgaggttg ccgacgttgg 960caaagtcttt actgttgcct ttccgtcacc tttagcagcc acgtccatct caatgcttgc 1020gccttcaggt tcagtcgttt accaacgccg catcttcggc ctccgaatta ttcagtatca 1080ttgacaaacc cacgcagctc gacccttctc gacccttttt ggaaagcagc cagagggctg 1140cttaggtcaa attgagatcc aaaacctggc atttgcctac ccctcccgac catctgccca 1200agtacttcga gatttcaact tgacaattcc agctggcaag acgacggccc tcgtcggtgc 1260atcaggtagc ggcaaaagca caatggtcgg cttacttgaa cggtggtatc tgcccagttc 1320ggggaggata ttacttgatg ggttggaact gggacaatac aatgtgaaat ggctgagaag 1380ccgcattcgc ctcgttcaac aggaacctgt gttgtttcgt ggcacaatct tccagaacat 1440tgccaacggt ttcatggatg agcaacgaga tctgcctcgc gaaaaacaaa tggagcttgt 1500gcaaaaagct tgcaaagcag caatgccgac gtgttcatta atgagcttcc gaacggttat 1560gagactgaag ttggcgagcg agccggagcc ttgagtggag gtcaacaagc cgaattgcaa 1620tcgcacgaag tatcatatcg gatcccaaga tcctgttact cgatgaagct accagcgccc 1680ttgacccgaa ggcggagaaa gtggtccagg aggccttgaa ccgagtgtcc aaagaccgca 1740ctactttggt cattgcccac aaactagcca ctgtcatacg actcactatt agggcgaatt 1800gggccctcta gatgcatgct cgagcggccg ccagtgtgac gaattgatgc agaattcggc 1860ttgtcattac gccgcactgg tgcgtgcaca ggacctcggg gctgacgaac aagaagaaca 1920tgagaagacc ctgcacgaaa aggcagcacg agaagctgct ggtgaacgac cggcacttga 1980gcgcactcac accactgcca catctcaagc tggagacctg gagaagcgga aggtgccggt 2040cgggactttg ggctactcgc tcctaaaatg catcctaatc atgttctacg aacaaaaaaa 2100tctctactgg tgcttcttgt tgtcaacaat agcggttctg atatgcgcgg ccacatttcc 2160aggacaagcc cttttgtttt cgagattgct cactgtcttc gagttgagtg gtcatgcggc 2220acaggaacgg gcagactttt atagtctgat gttctttgtc gtggctctag gaaatctagt 2280aggatatttc acgattggct ggacatgcaa cgttgtttca caagttgtca cccatcgcta 2340tcgagccgaa atgttccaac gagtactgga tcaagacatc gaattcttcg acatcccgga 2400gaatacttct ggtgctctca catcgcaact gtcagctcta cccacgcagt tgcaggagtt 2460gatatcaaca aattcttctc atttttatcg ttgtcgtaca acatcctctc gagcagtgct 2520ctagcactag cctatggatg gaaactgggc ctggtggttg tgtttggtgc acttccaccc 2580ctgcttttgg ctggctacct cagaattcgt cttgagacga agctagaagc cggaaactcg 2640gcaaactttg cagaaagtgc tgggcttgca agcgaagcag ttaccgcgat ccggaccgtc 2700tcatctttga ctctcgaagg scatgttctc caacagtact cggacatgtt gagcaaggtc 2760gtgctaagat catccaaagc tttggtttgg acgatgtttt ggttctcact gtcacagtcg 2820atcgagtttc tggctatggc cctgggaatt ttggtatggg aagtcgacta ctggcttcag 2880gtgaggtacg acacaactca attttatatc atcttcgtgg gcgttttgtt tgccggtcca 2940agcagcagcc cagaagccga attactccac gagtcttacc aaggctcggt cggctgcgaa 3000ctatatcctc tggctgcgga cattgaagcc gaccatccgc gaaacggagg agaacaagaa 3060aaaagggcca gtgggtggat gccctgtcga cctcgaggac attgaattca ggtatcgtca 3120acgtgattcg gctcgagttc tccgcggggt ttccatgaca atcgagccag gacaatttgt 3180agcttatgtg ggcgcttctg gctgtggcaa gtcaacgttg atcgctttgt tggaacgatt 3240ctacgacccg acctcgggcc gaatttcatt tgcacacgag aatattgcag aaatgtcgcc 3300gcgcttgtac cgcggccata tgtctttggt ccaacaggaa cccacayttt accaaggctc 3360cgttcgcgag aatgtgacgt tggccctcga agccgaatta tcagaagagc tttgtcaagg 3420acgccttccc gcaaggccaa tgctttggat tttgtcatct ctttaccaga aggctttgaa 3480acgccttgcg gctcaacgag ggatgcagtt ctccggcggg caacgacagc ggatcgccat 3540cgcaagagca ttgattcgaa atccaaagct gttgctactt gacgaagcga cgtcagccct 3600cgacacgcaa tcggaacgtc tggttcaagc tgccctcgat gaggcatcca cgagccgaac 3660gacaatagca gtggcgcacc gactttccac tattcggaat gttgatgtta tttttgtgtt 3720tgccaacggg agaatcgccg aaacgggcac tcacgcggaa ctacaacgac tgagaggaag 3780atattacgag atgtgtttgg cacaatcttt agaccaagca tgagcgttca cagagaagcg 3840gaaaagggcg gtgggatctt ttaggatagg tttagtggcg tgttacttac tacaggcgtt 3900tggattcagg tacgacaact tgtacaataa gtagcataga gcatgtaatg aaagggtact 3960cgtcccggaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaa 3999 10 3792 DNA Exophiala spinifera CDS (1)...(3792) misc_feature (0)...(0) p-glycoprotein, fully spliced cDNA 10atg gca gat gaa tcg gag aaa cct cga cca aac caa gat ggc agt gag 48Met Ala Asp Glu Ser Glu Lys Pro Arg Pro Asn Gln Asp Gly Ser Glu 1 5 10 15tcg tcc tca cac cct ccc cca gaa aag gaa acc gaa ggc agt att tca 96Ser Ser Ser His Pro Pro Pro Glu Lys Glu Thr Glu Gly Ser Ile Ser 20 25 30gac tat cta cga atc ttc aga tat gcc gac aaa tac gac tgg act ctc 144Asp Tyr Leu Arg Ile Phe Arg Tyr Ala Asp Lys Tyr Asp Trp Thr Leu 35 40 45aat gtc atc gcg ctc atc tgc gcc atc gga tcc ggg gct tcc ctt cct 192Asn Val Ile Ala Leu Ile Cys Ala Ile Gly Ser Gly Ala Ser Leu Pro 50 55 60ctg atg tcg atc atc ttc ggt agc ttc acc aac aag ttc aac aat tac 240Leu Met Ser Ile Ile Phe Gly Ser Phe Thr Asn Lys Phe Asn Asn Tyr 65 70 75 80aat tcg ggc gac ggg agt cct gaa gcg ttc aag gcc gat gtg gat cat 288Asn Ser Gly Asp Gly Ser Pro Glu Ala Phe Lys Ala Asp Val Asp His 85 90 95ttc gtc ctg tgg ttc gtc tac ctc ttt att ggg aag ttt gtc ctc acg 336Phe Val Leu Trp Phe Val Tyr Leu Phe Ile Gly Lys Phe Val Leu Thr 100 105 110tac gtt tcc acg gct gcc att acc att tca gct ata cga acc act cga 384Tyr Val Ser Thr Ala Ala Ile Thr Ile Ser Ala Ile Arg Thr Thr Arg 115 120 125act ctt cga cga gtg ttc ctt gaa tgc acc ttg cgg caa gag gtc tgg 432Thr Leu Arg Arg Val Phe Leu Glu Cys Thr Leu Arg Gln Glu Val Trp 130 135 140cat ttc gac aag cag agc aat gga gca atc gcc act car gtc act acc 480His Phe Asp Lys Gln Ser Asn Gly Ala Ile Ala Thr Gln Val Thr Thr145 150 155 160aat ggc aac cgt ata caa aca ggt att gcc gag aaa ttg gtc ttt acc 528Asn Gly Asn Arg Ile Gln Thr Gly Ile Ala Glu Lys Leu Val Phe Thr 165 170 175gtg cag gca ctt tca atg ttc ttt tct gca ttt gtg gtc gct ttg gcg 576Val Gln Ala Leu Ser Met Phe Phe Ser Ala Phe Val Val Ala Leu Ala 180 185 190tct cag tgg aag cta gct tta atc acc atg tcc gtc atc cct gcc att 624Ser Gln Trp Lys Leu Ala Leu Ile Thr Met Ser Val Ile Pro Ala Ile 195 200 205ttc ctg gtc acc ggc atc tgc ata gca att gat gcc gct cag gag gcc 672Phe Leu Val Thr Gly Ile Cys Ile Ala Ile Asp Ala Ala Gln Glu Ala 210 215 220agg atc acc agg atc tac tca cgc gcc gct gtc ctc gca gaa gaa gtc 720Arg Ile Thr Arg Ile Tyr Ser Arg Ala Ala Val Leu Ala Glu Glu Val225 230 235 240tta tca tcc atc cgg aca gtc cat gct ttc tac gcc cag aag aaa atg 768Leu Ser Ser Ile Arg Thr Val His Ala Phe Tyr Ala Gln Lys Lys Met 245 250 255gtc gaa aaa tat gat gtc ttt ttg cag caa gca cac caa gaa ggg aag 816Val Glu Lys Tyr Asp Val Phe Leu Gln Gln Ala His Gln Glu Gly Lys 260 265 270aag aaa tcg cca aat aat ggs gtc ttg ttc tca act gag tac ttt tgc 864Lys Lys Ser Pro Asn Asn Xaa Val Leu Phe Ser Thr Glu Tyr Phe Cys 275 280 285att tac gct gct atc gca ctg gcc ttt tgg aaa ggt ttt cgc atg tat 912Ile Tyr Ala Ala Ile Ala Leu Ala Phe Trp Lys Gly Phe Arg Met Tyr 290 295 300cag aat ggc gag gtt gcc gac gtt ggc aaa gtc ttt act gtt gtc ctt 960Gln Asn Gly Glu Val Ala Asp Val Gly Lys Val Phe Thr Val Val Leu305 310 315 320tcc gtc acc tta gca gcc acg tcc atc tca atg ctt gcg cct tca ggt 1008Ser Val Thr Leu Ala Ala Thr Ser Ile Ser Met Leu Ala Pro Ser Gly 325 330 335tca gtc gtt tac caa cgc cgc atc ttc ggc tcc gaa tta ttc agt atc 1056Ser Val Val Tyr Gln Arg Arg Ile Phe Gly Ser Glu Leu Phe Ser Ile 340 345 350att gac aaa ccc acg cag ctc gac cct ctc gac cct tct gga aag cag 1104Ile Asp Lys Pro Thr Gln Leu Asp Pro Leu Asp Pro Ser Gly Lys Gln 355 360 365cca gag ggc tgc cta ggt caa att gag atc caa aac ctg gca ttt gcc 1152Pro Glu Gly Cys Leu Gly Gln Ile Glu Ile Gln Asn Leu Ala Phe Ala 370 375 380tac ccc tcc cga cca tct gcc caa gta ctt cga gat ttc aac ttg aca 1200Tyr Pro Ser Arg Pro Ser Ala Gln Val Leu Arg Asp Phe Asn Leu Thr385 390 395 400att cca gct ggc aag acg acg gcc ctc gtc ggt gca tca ggt agc ggc 1248Ile Pro Ala Gly Lys Thr Thr Ala Leu Val Gly Ala Ser Gly Ser Gly 405 410 415aaa agc aca atg gtc ggc tta ctt gaa cgg tgg tat ctg ccc agt tcg 1296Lys Ser Thr Met Val Gly Leu Leu Glu Arg Trp Tyr Leu Pro Ser Ser 420 425 430ggg agg ata tta ctt gat ggg ttg gaa ctg gga caa tac aat gtg aaa 1344Gly Arg Ile Leu Leu Asp Gly Leu Glu Leu Gly Gln Tyr Asn Val Lys 435 440 445tgg ctg aga agc cgc att cgc ctc gtt caa cag gaa cct gtg ttg ttt 1392Trp Leu Arg Ser Arg Ile Arg Leu Val Gln Gln Glu Pro Val Leu Phe 450 455 460cgt ggc aca atc ttc cag aac att gcc aac ggt ttc atg gat gag caa 1440Arg Gly Thr Ile Phe Gln Asn Ile Ala Asn Gly Phe Met Asp Glu Gln465 470 475 480cga gat ctg cct cgc gaa aaa caa atg gag ctt gtg caa aaa gct tgc 1488Arg Asp Leu Pro Arg Glu Lys Gln Met Glu Leu Val Gln Lys Ala Cys 485 490 495aaa gcc agc aat ggc gac gtg ttc att aat gag ctt ccg aac ggt tat 1536Lys Ala Ser Asn Gly Asp Val Phe Ile Asn Glu Leu Pro Asn Gly Tyr 500 505 510gag act gaa gtt ggc gag cga gcc gga gcc ttg agt gga ggt caa cga 1584Glu Thr Glu Val Gly Glu Arg Ala Gly Ala Leu Ser Gly Gly Gln Arg 515 520 525caa cga att gca atc gca cga agt atc ata tcg gat ccc aag atc ctg 1632Gln Arg Ile Ala Ile Ala Arg Ser Ile Ile Ser Asp Pro Lys Ile Leu 530 535 540tta ctc gat gaa gct acc agc gcc ctt gac ccg aag gcg gag aaa gtg 1680Leu Leu Asp Glu Ala Thr Ser Ala Leu Asp Pro Lys Ala Glu Lys Val545 550 555 560gtc cag gag gcc ttg aac cga gtg tcc aaa gac cgc act act ttg gtc 1728Val Gln Glu Ala Leu Asn Arg Val Ser Lys Asp Arg Thr Thr Leu Val 565 570 575att gcc cac aaa cta gcc act gtc aaa agt gct ggc aac atc gca gtc 1776Ile Ala His Lys Leu Ala Thr Val Lys Ser Ala Gly Asn Ile Ala Val 580 585 590att tcc cag ggg aaa atc gtc gag caa ggc aca cac cac gaa ttg atc 1824Ile Ser Gln Gly Lys Ile Val Glu Gln Gly Thr His His Glu Leu Ile 595 600 605gaa ttc ggc tgt cat tac gcc gca ctg gtg cgt gca cag gac ctc ggg 1872Glu Phe Gly Cys His Tyr Ala Ala Leu Val Arg Ala Gln Asp Leu Gly 610 615 620gct gac gaa caa caa gaa cat gag aag acc ctg cac gaa aag gca gca 1920Ala Asp Glu Gln Gln Glu His Glu Lys Thr Leu His Glu Lys Ala Ala625 630 635 640cga gaa gct gct ggt gaa cga ccg gca ctt gag cgc act cac acc act 1968Arg Glu Ala Ala Gly Glu Arg Pro Ala Leu Glu Arg Thr His Thr Thr 645 650 655gcc aca tct caa gct gga gac ctg gag aag cgg aag gtg ccg gtc ggg 2016Ala Thr Ser Gln Ala Gly Asp Leu Glu Lys Arg Lys Val Pro Val Gly 660 665 670act ttg ggc tac tcg ctc cta aaa tgc atc cta atc atg ttc tac gaa 2064Thr Leu Gly Tyr Ser Leu Leu Lys Cys Ile Leu Ile Met Phe Tyr Glu 675 680 685caa aaa aat ctc tac tgg tgc ttc ttg ttg tca aca ata acg gtt ctg 2112Gln Lys Asn Leu Tyr Trp Cys Phe Leu Leu Ser Thr Ile Thr Val Leu 690 695 700ata tgc gcg gcc aca ttt cca gga caa gcc ctt ttg ttt tcg aga ttg 2160Ile Cys Ala Ala Thr Phe Pro Gly Gln Ala Leu Leu Phe Ser Arg Leu705 710 715 720ctc act gtc ttc gag ttg agt ggt cat gcg gca cag gaa cgg gca gac 2208Leu Thr Val Phe Glu Leu Ser Gly His Ala Ala Gln Glu Arg Ala Asp 725 730 735ttt tat att ctg atg ttc ttt gtc gtg gct cta gga aat cta gta gga 2256Phe Tyr Ile Leu Met Phe Phe Val Val Ala Leu Gly Asn Leu Val Gly 740 745 750tat ttc acg att ggc tgg aca tgc aac gtt att tca caa gtt gtc acc 2304Tyr Phe Thr Ile Gly Trp Thr Cys Asn Val Ile Ser Gln Val Val Thr 755 760 765cat cgc tat caa gcc gca atg ttc caa cga gta ctg gat caa gac atc 2352His Arg Tyr Gln Ala Ala Met Phe Gln Arg Val Leu Asp Gln Asp Ile 770 775 780gaa ctc ctc gac atc ccg gag caa att tct ggt gct ctc aca tcg caa 2400Glu Leu Leu Asp Ile Pro Glu Gln Ile Ser Gly Ala Leu Thr Ser Gln785 790 795 800ctg tca gct cta ccc acg cag ttg caa gag ttg ata tca gca aat ttt 2448Leu Ser Ala Leu Pro Thr Gln Leu Gln Glu Leu Ile Ser Ala Asn Phe 805 810 815ctc att tat atc gtt gtc ggt caa cat cgt ctc gag cag tgc tct acc 2496Leu Ile Tyr Ile Val Val Gly Gln His Arg Leu Glu Gln Cys Ser Thr 820 825 830act agc cta tgg atg gaa act ggg cct ggt ggt tgt gtt tgg tgc act 2544Thr Ser Leu Trp Met Glu Thr Gly Pro Gly Gly Cys Val Trp Cys Thr 835 840 845tcc acc cct gct ttt ggc tgg cta cct cag aat tcg tct aga gac gaa 2592Ser Thr Pro Ala Phe Gly Trp Leu Pro Gln Asn Ser Ser Arg Asp Glu 850 855 860gct aga agc cgg aaa ctc ggc aaa ctt tgc aga aag tgc tgg gct tgc 2640Ala Arg Ser Arg Lys Leu Gly Lys Leu Cys Arg Lys Cys Trp Ala Cys865 870 875 880aag cga agc agt tac cgc gat ccg gac cgt ctc atc ttt gac tct cga 2688Lys Arg Ser Ser Tyr Arg Asp Pro Asp Arg Leu Ile Phe Asp Ser Arg 885 890 895agg cca tgt tct cca aca gta ctc gga cat gtt gag caa ggt ctt gct 2736Arg Pro Cys Ser Pro Thr Val Leu Gly His Val Glu Gln Gly Leu Ala 900 905 910aag atc atc caa agc ttt tgg ttt gga cga tgt ttt ggt ttt cac ttg 2784Lys Ile Ile Gln Ser Phe Trp Phe Gly Arg Cys Phe Gly Phe His Leu 915 920 925tca cag tcg atg gag ttt ttg gct att gcc ctg gga ttt tgt att gca 2832Ser Gln Ser Met Glu Phe Leu Ala Ile Ala Leu Gly Phe Cys Ile Ala 930 935 940gtc gat aat tgg ctt cag gtg agt acg aca caa ctc aat ttt ata tca 2880Val Asp Asn Trp Leu Gln Val Ser Thr Thr Gln Leu Asn Phe Ile Ser945 950 955 960tct tcg tgg gcg ttt tgt ttg ccg gtc caa gca gca gcc cag tat ttg 2928Ser Ser Trp Ala Phe Cys Leu Pro Val Gln Ala Ala Ala Gln Tyr Leu 965 970 975gct tac tcc acg agt ttt acc aag gct cgg tcg gct gcg aac tat atc 2976Ala Tyr Ser Thr Ser Phe Thr Lys Ala Arg Ser Ala Ala Asn Tyr Ile 980 985 990ctc tgg ctg cgg aca ttg aag ccg acc atc cgc gaa acg gag gag aac 3024Leu Trp Leu Arg Thr Leu Lys Pro Thr Ile Arg Glu Thr Glu Glu Asn 995 1000 1005aag aaa aaa ggc cca gtg ggt gga tgc cct gtc gac ctc gag gac att 3072Lys Lys Lys Gly Pro Val Gly Gly Cys Pro Val Asp Leu Glu Asp Ile 1010 1015 1020gaa ttc agg tat cgt caa cgt gat tcg gct cga gtt ctc cgc ggg gtt 3120Glu Phe Arg Tyr Arg Gln Arg Asp Ser Ala Arg Val Leu Arg Gly Val1025 1030 1035 1040tcc atg aca atc gag cca gga caa ttt gta gct tat gtg ggc gct tct 3168Ser Met Thr Ile Glu Pro Gly Gln Phe Val Ala Tyr Val Gly Ala Ser 1045 1050 1055ggc tgt ggc aag tca acg ttg atc gct ttg tcg gaa cga ttc tac gac 3216Gly Cys Gly Lys Ser Thr Leu Ile Ala Leu Ser Glu Arg Phe Tyr Asp 1060 1065 1070ccg acc tcg ggc cga att tca ttt gca cac gag aat att gca gaa atg 3264Pro Thr Ser Gly Arg Ile Ser Phe Ala His Glu Asn Ile Ala Glu Met 1075 1080 1085tcg ccg cgc ttg tac cgc ggc cat atg tct ttg gtc caa cag gaa ccc 3312Ser Pro Arg Leu Tyr Arg Gly His Met Ser Leu Val Gln Gln Glu Pro 1090 1095 1100aca ctt tac caa ggc tcc gtt cgc gag aat gtg acg ttg gcc ctc gaa 3360Thr Leu Tyr Gln Gly Ser Val Arg Glu Asn Val Thr Leu Ala Leu Glu1105 1110 1115 1120gcc gaa tta tca gaa gag ctt tgt caa gga cgc ctt ccc gca agg cca 3408Ala Glu Leu Ser Glu Glu Leu Cys Gln Gly Arg Leu Pro Ala Arg Pro 1125 1130 1135atg ctt tgg att ttg tca tct ctt tac cag aag gct ttg aaa cgc ctt 3456Met Leu Trp Ile Leu Ser Ser Leu Tyr Gln Lys Ala Leu Lys Arg Leu 1140 1145 1150gcg gct caa cga ggg atg cag ttc tcc ggc ggg caa cga cag cgg atc 3504Ala Ala Gln Arg Gly Met Gln Phe Ser Gly Gly Gln Arg Gln Arg Ile 1155 1160 1165gcc atc gca aga gca ttg att cga aat cca aag ctg ttg cta ctt gac 3552Ala Ile Ala Arg Ala Leu Ile Arg Asn Pro Lys Leu Leu Leu Leu Asp 1170 1175 1180gaa gcg acg tca gcc ctc gac acg caa tcg gaa cgt ctg gtt caa gct 3600Glu Ala Thr Ser Ala Leu Asp Thr Gln Ser Glu Arg Leu Val Gln Ala1185 1190 1195 1200gcc ctc gat gag gca tcc acg agc cga acg aca ata gca gtg gcg cac 3648Ala Leu Asp Glu Ala Ser Thr Ser Arg Thr Thr Ile Ala Val Ala His 1205 1210 1215cga ctt tcc act att cgg aat gtt gat gtt att ttt gtg ttt gcc aac 3696Arg Leu Ser Thr Ile Arg Asn Val Asp Val Ile Phe Val Phe Ala Asn 1220 1225 1230ggg aga atc gcc gaa acg ggc act cac gcg gaa cta caa cga ctg aga 3744Gly Arg Ile Ala Glu Thr Gly Thr His Ala Glu Leu Gln Arg Leu Arg 1235 1240 1245gga aga tat tac gag atg tgt ttg gca caa tct tta gac caa gca tga 3792Gly Arg Tyr Tyr Glu Met Cys Leu Ala Gln Ser Leu Asp Gln Ala * 1250 1255 1260 11 1263 PRT Exophiala spinifera UNSURE (0)...(0) Xaa is any amino acid 11Met Ala Asp Glu Ser Glu Lys Pro Arg Pro Asn Gln Asp Gly Ser Glu 1 5 10 15Ser Ser Ser His Pro Pro Pro Glu Lys Glu Thr Glu Gly Ser Ile Ser 20 25 30Asp Tyr Leu Arg Ile Phe Arg Tyr Ala Asp Lys Tyr Asp Trp Thr Leu 35 40 45Asn Val Ile Ala Leu Ile Cys Ala Ile Gly Ser Gly Ala Ser Leu Pro 50 55 60Leu Met Ser Ile Ile Phe Gly Ser Phe Thr Asn Lys Phe Asn Asn Tyr65 70 75 80Asn Ser Gly Asp Gly Ser Pro Glu Ala Phe Lys Ala Asp Val Asp His 85 90 95Phe Val Leu Trp Phe Val Tyr Leu Phe Ile Gly Lys Phe Val Leu Thr 100 105 110Tyr Val Ser Thr Ala Ala Ile Thr Ile Ser Ala Ile Arg Thr Thr Arg 115 120 125Thr Leu Arg Arg Val Phe Leu Glu Cys Thr Leu Arg Gln Glu Val Trp 130 135 140His Phe Asp Lys Gln Ser Asn Gly Ala Ile Ala Thr Gln Val Thr Thr145 150 155 160Asn Gly Asn Arg Ile Gln Thr Gly Ile Ala Glu Lys Leu Val Phe Thr 165 170 175Val Gln Ala Leu Ser Met Phe Phe Ser Ala Phe Val Val Ala Leu Ala 180 185 190Ser Gln Trp Lys Leu Ala Leu Ile Thr Met Ser Val Ile Pro Ala Ile 195 200 205Phe Leu Val Thr Gly Ile Cys Ile Ala Ile Asp Ala Ala Gln Glu Ala 210 215 220Arg Ile Thr Arg Ile Tyr Ser Arg Ala Ala Val Leu Ala Glu Glu Val225 230 235 240Leu Ser Ser Ile Arg Thr Val His Ala Phe Tyr Ala Gln Lys Lys Met 245 250 255Val Glu Lys Tyr Asp Val Phe Leu Gln Gln Ala His Gln Glu Gly Lys 260 265 270Lys Lys Ser Pro Asn Asn Xaa Val Leu Phe Ser Thr Glu Tyr Phe Cys 275 280 285Ile Tyr Ala Ala Ile Ala Leu Ala Phe Trp Lys Gly Phe Arg Met Tyr 290 295 300Gln Asn Gly Glu Val Ala Asp Val Gly Lys Val Phe Thr Val Val Leu305 310 315 320Ser Val Thr Leu Ala Ala Thr Ser Ile Ser Met Leu Ala Pro Ser Gly 325 330 335Ser Val Val Tyr Gln Arg Arg Ile Phe Gly Ser Glu Leu Phe Ser Ile 340 345 350Ile Asp Lys Pro Thr Gln Leu Asp Pro Leu Asp Pro Ser Gly Lys Gln 355 360 365Pro Glu Gly Cys Leu Gly Gln Ile Glu Ile Gln Asn Leu Ala Phe Ala 370 375 380Tyr Pro Ser Arg Pro Ser Ala Gln Val Leu Arg Asp Phe Asn Leu Thr385 390 395 400Ile Pro Ala Gly Lys Thr Thr Ala Leu Val Gly Ala Ser Gly Ser Gly 405 410 415Lys Ser Thr Met Val Gly Leu Leu Glu Arg Trp Tyr Leu Pro Ser Ser 420 425 430Gly Arg Ile Leu Leu Asp Gly Leu Glu Leu Gly Gln Tyr Asn Val Lys 435 440 445Trp Leu Arg Ser Arg Ile Arg Leu Val Gln Gln Glu Pro Val Leu Phe 450 455 460Arg Gly Thr Ile Phe Gln Asn Ile Ala Asn Gly Phe Met Asp Glu Gln465 470 475 480Arg Asp Leu Pro Arg Glu Lys Gln Met Glu Leu Val Gln Lys Ala Cys 485 490 495Lys Ala Ser Asn Gly Asp Val Phe Ile Asn Glu Leu Pro Asn Gly Tyr 500 505 510Glu Thr Glu Val Gly Glu Arg Ala Gly Ala Leu Ser Gly Gly Gln Arg 515 520 525Gln Arg Ile Ala Ile Ala Arg Ser Ile Ile Ser Asp Pro Lys Ile Leu 530 535 540Leu Leu Asp Glu Ala Thr Ser Ala Leu Asp Pro Lys Ala Glu Lys Val545 550 555 560Val Gln Glu Ala Leu Asn Arg Val Ser Lys Asp Arg Thr Thr Leu Val 565 570 575Ile Ala His Lys Leu Ala Thr Val Lys Ser Ala Gly Asn Ile Ala Val 580 585 590Ile Ser Gln Gly Lys Ile Val Glu Gln Gly Thr His His Glu Leu Ile 595 600 605Glu Phe Gly Cys His Tyr Ala Ala Leu Val Arg Ala Gln Asp Leu Gly 610 615 620Ala Asp Glu Gln Gln Glu His Glu Lys Thr Leu His Glu Lys Ala Ala625 630 635 640Arg Glu Ala Ala Gly Glu Arg Pro Ala Leu Glu Arg Thr His Thr Thr 645 650 655Ala Thr Ser Gln Ala Gly Asp Leu Glu Lys Arg Lys Val Pro Val Gly 660 665 670Thr Leu Gly Tyr Ser Leu Leu Lys Cys Ile Leu Ile Met Phe Tyr Glu 675 680 685Gln Lys Asn Leu Tyr Trp Cys Phe Leu Leu Ser Thr Ile Thr Val Leu 690 695 700Ile Cys Ala Ala Thr Phe Pro Gly Gln Ala Leu Leu Phe Ser Arg Leu705 710 715 720Leu Thr Val Phe Glu Leu Ser Gly His Ala Ala Gln Glu Arg Ala Asp 725 730 735Phe Tyr Ile Leu Met Phe Phe Val Val Ala Leu Gly Asn Leu Val Gly 740 745 750Tyr Phe Thr Ile Gly Trp Thr Cys Asn Val Ile Ser Gln Val Val Thr 755 760 765His Arg Tyr Gln Ala Ala Met Phe Gln Arg Val Leu Asp Gln Asp Ile 770 775 780Glu Leu Leu Asp Ile Pro Glu Gln Ile Ser Gly Ala Leu Thr Ser Gln785 790 795 800Leu Ser Ala Leu Pro Thr Gln Leu Gln Glu Leu Ile Ser Ala Asn Phe 805 810 815Leu Ile Tyr Ile Val Val Gly Gln His Arg Leu Glu Gln Cys Ser Thr 820 825 830Thr Ser Leu Trp Met Glu Thr Gly Pro Gly Gly Cys Val Trp Cys Thr 835 840 845Ser Thr Pro Ala Phe Gly Trp Leu Pro Gln Asn Ser Ser Arg Asp Glu 850 855 860Ala Arg Ser Arg Lys Leu Gly Lys Leu Cys Arg Lys Cys Trp Ala Cys865 870 875 880Lys Arg Ser Ser Tyr Arg Asp Pro Asp Arg Leu Ile Phe Asp Ser Arg 885 890 895Arg Pro Cys Ser Pro Thr Val Leu Gly His Val Glu Gln Gly Leu Ala 900 905 910Lys Ile Ile Gln Ser Phe Trp Phe Gly Arg Cys Phe Gly Phe His Leu 915 920 925Ser Gln Ser Met Glu Phe Leu Ala Ile Ala Leu Gly Phe Cys Ile Ala 930 935 940Val Asp Asn Trp Leu Gln Val Ser Thr Thr Gln Leu Asn Phe Ile Ser945 950 955 960Ser Ser Trp Ala Phe Cys Leu Pro Val Gln Ala Ala Ala Gln Tyr Leu 965 970 975Ala Tyr Ser Thr Ser Phe Thr Lys Ala Arg Ser Ala Ala Asn Tyr Ile 980 985 990Leu Trp Leu Arg Thr Leu Lys Pro Thr Ile Arg Glu Thr Glu Glu Asn 995 1000 1005Lys Lys Lys Gly Pro Val Gly Gly Cys Pro Val Asp Leu Glu Asp Ile 1010 1015 1020Glu Phe Arg Tyr Arg Gln Arg Asp Ser Ala Arg Val Leu Arg Gly Val1025 1030 1035 1040Ser Met Thr Ile Glu Pro Gly Gln Phe Val Ala Tyr Val Gly Ala Ser 1045 1050 1055Gly Cys Gly Lys Ser Thr Leu Ile Ala Leu Ser Glu Arg Phe Tyr Asp 1060 1065 1070Pro Thr Ser Gly Arg Ile Ser Phe Ala His Glu Asn Ile Ala Glu Met 1075 1080 1085Ser Pro Arg Leu Tyr Arg Gly His Met Ser Leu Val Gln Gln Glu Pro 1090 1095 1100Thr Leu Tyr Gln Gly Ser Val Arg Glu Asn Val Thr Leu Ala Leu Glu1105 1110 1115 1120Ala Glu Leu Ser Glu Glu Leu Cys Gln Gly Arg Leu Pro Ala Arg Pro 1125 1130 1135Met Leu Trp Ile Leu Ser Ser Leu Tyr Gln Lys Ala Leu Lys Arg Leu 1140 1145 1150Ala Ala Gln Arg Gly Met Gln Phe Ser Gly Gly Gln Arg Gln Arg Ile 1155 1160 1165Ala Ile Ala Arg Ala Leu Ile Arg Asn Pro Lys Leu Leu Leu Leu Asp 1170 1175 1180Glu Ala Thr Ser Ala Leu Asp Thr Gln Ser Glu Arg Leu Val Gln Ala1185 1190 1195 1200Ala Leu Asp Glu Ala Ser Thr Ser Arg Thr Thr Ile Ala Val Ala His 1205 1210 1215Arg Leu Ser Thr Ile Arg Asn Val Asp Val Ile Phe Val Phe Ala Asn 1220 1225 1230Gly Arg Ile Ala Glu Thr Gly Thr His Ala Glu Leu Gln Arg Leu Arg 1235 1240 1245Gly Arg Tyr Tyr Glu Met Cys Leu Ala Gln Ser Leu Asp Gln Ala 1250 1255 1260 12 1937 DNA Exophiala spinifera CDS (153)...(1736) 12gcggatccgt tttttttttt ttttttccta agttcgacta cccacttgct agtctcacag 60tagctccaag ggtataagtt cgactcgaag ctgcatctct ccgtgaaaca tggcaatagt 120ttttgtagac agatccatca accgagtaca cg atg ccg tca agg tac att ctc 173 Met Pro Ser Arg Tyr Ile Leu 1 5tct tgg ctc ctc acc tgc ttt ttg ggc att gct ttt ggc tca cga tgc 221Ser Trp Leu Leu Thr Cys Phe Leu Gly Ile Ala Phe Gly Ser Arg Cys 10 15 20ggg tcg tct gct cct act gtc aag att gat gct ggg atg gtg gtc ggc 269Gly Ser Ser Ala Pro Thr Val Lys Ile Asp Ala Gly Met Val Val Gly 25 30 35acg act act act gtc ccc ggc acc act gcg acc gtc agc gag ttc ttg 317Thr Thr Thr Thr Val Pro Gly Thr Thr Ala Thr Val Ser Glu Phe Leu 40 45 50 55ggc gtt cct ttt gcc gcc tct ccg aca cga ttt gcg cct cct act cgt 365Gly Val Pro Phe Ala Ala Ser Pro Thr Arg Phe Ala Pro Pro Thr Arg 60 65 70ccc gtg cct tgg tca acg cct ttg caa gcc act gca tat ggt cca gca 413Pro Val Pro Trp Ser Thr Pro Leu Gln Ala Thr Ala Tyr Gly Pro Ala 75 80 85tgc cct caa caa ttc aat tac ccc gaa gaa ctc cgt gag att acg atg 461Cys Pro Gln Gln Phe Asn Tyr Pro Glu Glu Leu Arg Glu Ile Thr Met 90 95 100gcc tgg ttc aat aca ccg ccc ccg tca gct ggt gaa agt gag gac tgc 509Ala Trp Phe Asn Thr Pro Pro Pro Ser Ala Gly Glu Ser Glu Asp Cys 105 110 115ctg aac ctc aac atc tac gtc cca gga act gag aac aca aac aaa gcc 557Leu Asn Leu Asn Ile Tyr Val Pro Gly Thr Glu Asn Thr Asn Lys Ala120 125 130 135gtc atg gtt tgg ata tac ggt gga gcg ctg gaa tat ggt tgg aat tca 605Val Met Val Trp Ile Tyr Gly Gly Ala Leu Glu Tyr Gly Trp Asn Ser 140 145 150ttc cac ctt tac gac ggg gct agt ttc gca gcc aat cag gat gtc atc 653Phe His Leu Tyr Asp Gly Ala Ser Phe Ala Ala Asn Gln Asp Val Ile 155 160 165gtc gtg acc atc aac tac aga acg aac att ctg ggg ttc cct gct gcc 701Val Val Thr Ile Asn Tyr Arg Thr Asn Ile Leu Gly Phe Pro Ala Ala 170 175 180cct cag ctt cca ata aca cag cga aat ctg ggg ttc cta gac caa agg 749Pro Gln Leu Pro Ile Thr Gln Arg Asn Leu Gly Phe Leu Asp Gln Arg 185 190 195ttt gct ttg gat tgg gta cag cgg aac atc gca gcc ttt ggc ggt gat 797Phe Ala Leu Asp Trp Val Gln Arg Asn Ile Ala Ala Phe Gly Gly Asp200 205 210 215cct cga aag gtc aca ata ttt ggg cag agt gcg ggg ggc aga agt gtc 845Pro Arg Lys Val Thr Ile Phe Gly Gln Ser Ala Gly Gly Arg Ser Val 220 225 230gac gtc ctc ttg acg tct atg cca cac aac cca ccc ttc cga gca gca 893Asp Val Leu Leu Thr Ser Met Pro His Asn Pro Pro Phe Arg Ala Ala 235 240 245atc atg gag tcc ggt gtg gct aac tac aac ttc ccc aag gga gat ttg 941Ile Met Glu Ser Gly Val Ala Asn Tyr Asn Phe Pro Lys Gly Asp Leu 250 255 260tcc gaa cct tgg aac acc act gtt caa gct ctc aac tgt acc acc agt 989Ser Glu Pro Trp Asn Thr Thr Val Gln Ala Leu Asn Cys Thr Thr Ser 265 270 275atc gac atc ttg agt tgt atg aga aga gtc gat ctc gcc act ctg atg 1037Ile Asp Ile Leu Ser Cys Met Arg Arg Val Asp Leu Ala Thr Leu Met280 285 290 295aac acg atc gag caa ctc gga ctt ggg ttt gag tac acg ttg gac aac 1085Asn Thr Ile Glu Gln Leu Gly Leu Gly Phe Glu Tyr Thr Leu Asp Asn 300 305 310gta acg gtt gtg tac cgt tct gaa acg gct cgc acg act ggt gac att 1133Val Thr Val Val Tyr Arg Ser Glu Thr Ala Arg Thr Thr Gly Asp Ile 315 320 325gct cgt gta cct gtt ctc gtc ggg acg gtg gcc aac gac gga ctt ctc 1181Ala Arg Val Pro Val Leu Val Gly Thr Val Ala Asn Asp Gly Leu Leu 330 335 340ttt gtc ctc ggg gag aat gac acc caa gca tat ctc gag gag gca atc 1229Phe Val Leu Gly Glu Asn Asp Thr Gln Ala Tyr Leu Glu Glu Ala Ile 345 350 355ccg aat cag ccc gac ctt tac cag act ctc ctt gga gca tat ccc att 1277Pro Asn Gln Pro Asp Leu Tyr Gln Thr Leu Leu Gly Ala Tyr Pro Ile360 365 370 375gga tcc cca ggg atc gga tcg cct caa gat cag att gcc gcc att gag 1325Gly Ser Pro Gly Ile Gly Ser Pro Gln Asp Gln Ile Ala Ala Ile Glu 380 385 390acc gag gta aga ttc cag tgt cct tct gcc atc gtg gct cag gac tcc 1373Thr Glu Val Arg Phe Gln Cys Pro Ser Ala Ile Val Ala Gln Asp Ser 395 400 405cgg aat cgg ggt atc cct tct tgg cgc tac tac tac aat gcg acc ttt 1421Arg Asn Arg Gly Ile Pro Ser Trp Arg Tyr Tyr Tyr Asn Ala Thr Phe 410 415 420gag aat ctg gag ctt ttc cct ggg tcc gaa gtg tac cac agc tct gaa 1469Glu Asn Leu Glu Leu Phe Pro Gly Ser Glu Val Tyr His Ser Ser Glu 425 430 435gtc ggg atg gtg ttt ggc acg tat cct gtc gca agt gcg acc gcc ttg 1517Val Gly Met Val Phe Gly Thr Tyr Pro Val Ala Ser Ala Thr Ala Leu440 445 450 455gag gcc cag acg agc aaa tac atg cag ggt gcc tgg gcg gcc ttt gcc 1565Glu Ala Gln Thr Ser Lys Tyr Met Gln Gly Ala Trp Ala Ala Phe Ala 460 465 470aaa aac ccc atg aat ggg cct ggg tgg aaa caa gtg ccg aat gtc gcg 1613Lys Asn Pro Met Asn Gly Pro Gly Trp Lys Gln Val Pro Asn Val Ala 475 480 485gcg ctt ggc tca cca ggc aaa gcc atc cag gtt gac gtc tct cca gcg 1661Ala Leu Gly Ser Pro Gly Lys Ala Ile Gln Val Asp Val Ser Pro Ala 490 495 500aca ata gac caa cga tgt gcc ttg tac acg cat tat tat act gag ttg 1709Thr Ile Asp Gln Arg Cys Ala Leu Tyr Thr His Tyr Tyr Thr Glu Leu 505 510 515ggc aca atc gcg ccg agg aca ttt tga ggaccagggt attgtaccta 1756Gly Thr Ile Ala Pro Arg Thr Phe *520 525cagcgggttc ggaaaaggag gtatctgctg tcaatttgcc gccagccatc attgaagagt 1816gctgaaattt catgggggaa tatccatcca tgctcacatt agcgcttttg gaagatggac 1876tgttagcgag tcttgggcgg tttcaggctt ttcccccccc aaaaaaaaaa aaaaaaaaaa 1936a 1937 13 527 PRT Exophiala spinifera 13Met Pro Ser Arg Tyr Ile Leu Ser Trp Leu Leu Thr Cys Phe Leu Gly 1 5 10 15Ile Ala Phe Gly Ser Arg Cys Gly Ser Ser Ala Pro Thr Val Lys Ile 20 25 30Asp Ala Gly Met Val Val Gly Thr Thr Thr Thr Val Pro Gly Thr Thr 35 40 45Ala Thr Val Ser Glu Phe Leu Gly Val Pro Phe Ala Ala Ser Pro Thr 50 55 60Arg Phe Ala Pro Pro Thr Arg Pro Val Pro Trp Ser Thr Pro Leu Gln65 70 75 80Ala Thr Ala Tyr Gly Pro Ala Cys Pro Gln Gln Phe Asn Tyr Pro Glu 85 90 95Glu Leu Arg Glu Ile Thr Met Ala Trp Phe Asn Thr Pro Pro Pro Ser 100 105 110Ala Gly Glu Ser Glu Asp Cys Leu Asn Leu Asn Ile Tyr Val Pro Gly 115 120 125Thr Glu Asn Thr Asn Lys Ala Val Met Val Trp Ile Tyr Gly Gly Ala 130 135 140Leu Glu Tyr Gly Trp Asn Ser Phe His Leu Tyr Asp Gly Ala Ser Phe145 150 155 160Ala Ala Asn Gln Asp Val Ile Val Val Thr Ile Asn Tyr Arg Thr Asn 165 170 175Ile Leu Gly Phe Pro Ala Ala Pro Gln Leu Pro Ile Thr Gln Arg Asn 180 185 190Leu Gly Phe Leu Asp Gln Arg Phe Ala Leu Asp Trp Val Gln Arg Asn 195 200 205Ile Ala Ala Phe Gly Gly Asp Pro Arg Lys Val Thr Ile Phe Gly Gln 210 215 220Ser Ala Gly Gly Arg Ser Val Asp Val Leu Leu Thr Ser Met Pro His225 230 235 240Asn Pro Pro Phe Arg Ala Ala Ile Met Glu Ser Gly Val Ala Asn Tyr 245 250 255Asn Phe Pro Lys Gly Asp Leu Ser Glu Pro Trp Asn Thr Thr Val Gln 260 265 270Ala Leu Asn Cys Thr Thr Ser Ile Asp Ile Leu Ser Cys Met Arg Arg 275 280 285Val Asp Leu Ala Thr Leu Met Asn Thr Ile Glu Gln Leu Gly Leu Gly 290 295 300Phe Glu Tyr Thr Leu Asp Asn Val Thr Val Val Tyr Arg Ser Glu Thr305 310 315 320Ala Arg Thr Thr Gly Asp Ile Ala Arg Val Pro Val Leu Val Gly Thr 325 330 335Val Ala Asn Asp Gly Leu Leu Phe Val Leu Gly Glu Asn Asp Thr Gln 340 345 350Ala Tyr Leu Glu Glu Ala Ile Pro Asn Gln Pro Asp Leu Tyr Gln Thr 355 360 365Leu Leu Gly Ala Tyr Pro Ile Gly Ser Pro Gly Ile Gly Ser Pro Gln 370 375 380Asp Gln Ile Ala Ala Ile Glu Thr Glu Val Arg Phe Gln Cys Pro Ser385 390 395 400Ala Ile Val Ala Gln Asp Ser Arg Asn Arg Gly Ile Pro Ser Trp Arg 405 410 415Tyr Tyr Tyr Asn Ala Thr Phe Glu Asn Leu Glu Leu Phe Pro Gly Ser 420 425 430Glu Val Tyr His Ser Ser Glu Val Gly Met Val Phe Gly Thr Tyr Pro 435 440 445Val Ala Ser Ala Thr Ala Leu Glu Ala Gln Thr Ser Lys Tyr Met Gln 450 455 460Gly Ala Trp Ala Ala Phe Ala Lys Asn Pro Met Asn Gly Pro Gly Trp465 470 475 480Lys Gln Val Pro Asn Val Ala Ala Leu Gly Ser Pro Gly Lys Ala Ile 485 490 495Gln Val Asp Val Ser Pro Ala Thr Ile Asp Gln Arg Cys Ala Leu Tyr 500 505 510Thr His Tyr Tyr Thr Glu Leu Gly Thr Ile Ala Pro Arg Thr Phe 515 520 525 14 1800 DNA Bacterium of ATCC 55552 CDS (94)...(1683) 14actagtggat cattgcattg gctggcggac tggcgcgccg atagtcgttg cgatggtcgc 60gagaataagc gtgcgaagtg ggaggatgtg aag atg ggg gcc agg agt atg tgt 114 Met Gly Ala Arg Ser Met Cys 1 5gcg gga cgg ttc gga cgc ttc tgc att ggc ttg gct tca tcg gtt gcc 162Ala Gly Arg Phe Gly Arg Phe Cys Ile Gly Leu Ala Ser Ser Val Ala 10 15 20gtg act cta ggg gga gcc tcc gcc gcc ggc gcg gca acc gcg acg gat 210Val Thr Leu Gly Gly Ala Ser Ala Ala Gly Ala Ala Thr Ala Thr Asp 25 30 35ttt ccg gtc cgc agg acc gat ctg ggc cag gtt cag gga ctg gcc ggg 258Phe Pro Val Arg Arg Thr Asp Leu Gly Gln Val Gln Gly Leu Ala Gly 40 45 50 55gac gtg atg agc ttt cgc gga ata ccc tat gca gcg ccg ccg gtg ggc 306Asp Val Met Ser Phe Arg Gly Ile Pro Tyr Ala Ala Pro Pro Val Gly 60 65 70ggg ctg cgt tgg aag ccg ccc caa cac gcc cgg ccc tgg gcg ggc gtt 354Gly Leu Arg Trp Lys Pro Pro Gln His Ala Arg Pro Trp Ala Gly Val 75 80 85cgc ccc gcc acc caa ttt ggc tcc gac tgc ttc ggc gcg gcc tat ctt 402Arg Pro Ala Thr Gln Phe Gly Ser Asp Cys Phe Gly Ala Ala Tyr Leu 90 95 100cgc aaa ggc agc ctc gcc ccc ggc gtg agc gag gac tgt ctt tac ctc 450Arg Lys Gly Ser Leu Ala Pro Gly Val Ser Glu Asp Cys Leu Tyr Leu 105 110 115aac gta tgg gcg ccg tca ggc gct aaa ccc ggc cag tac ccc gtc atg 498Asn Val Trp Ala Pro Ser Gly Ala Lys Pro Gly Gln Tyr Pro Val Met120 125 130 135gtc tgg gtc tac ggc ggc ggc ttc gcc ggc ggc acg gcc gcc atg ccc 546Val Trp Val Tyr Gly Gly Gly Phe Ala Gly Gly Thr Ala Ala Met Pro 140 145 150tac tac gac ggc gag gcg ctt gcg cga cag ggc gtc gtc gtg gtg acg 594Tyr Tyr Asp Gly Glu Ala Leu Ala Arg Gln Gly Val Val Val Val Thr 155 160 165ttt aac tat cgg acg aac atc ctg ggc ttt ttc gcc cat cct ggt ctc 642Phe Asn Tyr Arg Thr Asn Ile Leu Gly Phe Phe Ala His Pro Gly Leu 170 175 180tcg cgc gag agc ccc acc gga act tcg ggc aac tac ggc cta ctc gac 690Ser Arg Glu Ser Pro Thr Gly Thr Ser Gly Asn Tyr Gly Leu Leu Asp 185 190 195att ctc gcc gct ctt cgg tgg gtg cag agc aac gcc cgc gcc ttc gga 738Ile Leu Ala Ala Leu Arg Trp Val Gln Ser Asn Ala Arg Ala Phe Gly200 205 210 215ggg gac ccc ggc cga gtg acg gtc ttt ggt gaa tcg gcc gga gcg agc 786Gly Asp Pro Gly Arg Val Thr Val Phe Gly Glu Ser Ala Gly Ala Ser 220 225 230gcg atc gga ctt ctg ctc acc tcg ccg ctg agc aag ggt ctc ttc cgt 834Ala Ile Gly Leu Leu Leu Thr Ser Pro Leu Ser Lys Gly Leu Phe Arg 235 240 245ggc gct atc ctc gaa agt cca ggg ctg acg cga ccg ctc gcg acg ctc 882Gly Ala Ile Leu Glu Ser Pro Gly Leu Thr Arg Pro Leu Ala Thr Leu 250 255 260gcc gac agc gcc gcc tcg ggc gag cgc ctc gac gcc gat ctt tcg cga 930Ala Asp Ser Ala Ala Ser Gly Glu Arg Leu Asp Ala Asp Leu Ser Arg 265 270 275ctg cgc tcg acc gac cca gcc acc ctg atg gcg cgc gcc gac gcg gcc 978Leu Arg Ser Thr Asp Pro Ala Thr Leu Met Ala Arg Ala Asp Ala Ala280 285 290 295cgc ccg gca tcg cgg gac ctg cgc agg ccg cgt ccg acc gga ccg atc 1026Arg Pro Ala Ser Arg Asp Leu Arg Arg Pro Arg Pro Thr Gly Pro Ile 300 305 310gtc gat ggc cat gtg ctg ccg cag acc gac agc gcg gcg atc gcg gcg 1074Val Asp Gly His Val Leu Pro Gln Thr Asp Ser Ala Ala Ile Ala Ala 315 320 325ggg cag ctg gcg ccg gtt cgg gtc ctg atc gga acc aat gcc gac gaa 1122Gly Gln Leu Ala Pro Val Arg Val Leu Ile Gly Thr Asn Ala Asp Glu 330 335 340ggc cgc gcc ttc ctc ggg cgc gcg ccg atg gag acg cca gcg gac tac 1170Gly Arg Ala Phe Leu Gly Arg Ala Pro Met Glu Thr Pro Ala Asp Tyr 345 350 355caa gcc tat ctg gag gcg cag ttt ggc gac caa gcc gcc gcc gtg gcg 1218Gln Ala Tyr Leu Glu Ala Gln Phe Gly Asp Gln Ala Ala Ala Val Ala360 365 370 375gcg tgc tat ccc ctc gac ggc cgg gcc acg ccc aag gaa atg gtc gcg 1266Ala Cys Tyr Pro Leu Asp Gly Arg Ala Thr Pro Lys Glu Met Val Ala 380 385 390cgc atc ttc ggc gac aat cag ttc aat cgg ggg gtc tcg gcc ttc tcg 1314Arg Ile Phe Gly Asp Asn Gln Phe Asn Arg Gly Val Ser Ala Phe Ser 395 400 405gaa gcg ctt gtg cgc cag ggc gcg ccc gtg tgg cgt tat cag ttc aac 1362Glu Ala Leu Val Arg Gln Gly Ala Pro Val Trp Arg Tyr Gln Phe Asn 410 415 420ggt aat acc gag ggt gga aga gcg ccg gct acc cac gga gcc gaa att 1410Gly Asn Thr Glu Gly Gly Arg Ala Pro Ala Thr His Gly Ala Glu Ile 425 430 435ccc tac gtt ttc ggg gtg ttc aag ctc gac gag ttg ggt ctg ttc gat 1458Pro Tyr Val Phe Gly Val Phe Lys Leu Asp Glu Leu Gly Leu Phe Asp440 445 450 455tgg ccg ccc gag ggg ccc acg ccc gcc gac cgt gcg ctg ggc caa ctg 1506Trp Pro Pro Glu Gly Pro Thr Pro Ala Asp Arg Ala Leu Gly Gln Leu 460 465 470atg tcc tcc gcc tgg gtc cgg ttc gcc aag aat ggc gac ccc gcc ggg 1554Met Ser Ser Ala Trp Val Arg Phe Ala Lys Asn Gly Asp Pro Ala Gly 475 480 485gac gcc ctt acc tgg cct gcc tat tct acg ggc aag tcg acc atg aca 1602Asp Ala Leu Thr Trp Pro Ala Tyr Ser Thr Gly Lys Ser Thr Met Thr 490 495 500ttc ggt ccc gag ggc cgc gcg gcg gtg gtg tcg ccc gga cct tcc atc 1650Phe Gly Pro Glu Gly Arg Ala Ala Val Val Ser Pro Gly Pro Ser Ile 505 510 515ccc cct tgc gcg gat ggc gcc aag gcg ggg tga cgccgtcgac gatggcgtga 1703Pro Pro Cys Ala Asp Gly Ala Lys Ala Gly *520 525cgacggtcga ggcgatgttc tcgatctgga gtccgcgccg cctcgatttg cgtcgtctcc 1763ggcgctcaga cgaacgcccc agttccatcc acacagt 1800 15 529 PRT Bacterium of ATCC 55552 15Met 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 His65 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 16 1389 DNA Exophiala spinifera CDS (1)...(1389) misc_feature (0)...(0) truncated APAO 16gac aac gtt gcg gac gtg gta gtg gtg ggc gct ggc ttg agc ggt ttg 48Asp Asn Val Ala Asp Val Val Val Val Gly Ala Gly Leu Ser Gly Leu 1 5 10 15gag acg gca cgc aaa gtc cag gcc gcc ggt ctg tcc tgc ctc gtt ctt 96Glu Thr Ala Arg Lys Val Gln Ala Ala Gly Leu Ser Cys Leu Val Leu 20 25 30gag gcg atg gat cgt gta ggg gga aag act ctg agc gta caa tcg ggt 144Glu Ala Met Asp Arg Val Gly Gly Lys Thr Leu Ser Val Gln Ser Gly 35 40 45ccc ggc agg acg act atc aac gac ctc ggc gct gcg tgg atc aat gac 192Pro Gly Arg Thr Thr Ile Asn Asp Leu Gly Ala Ala Trp Ile Asn Asp 50 55 60agc aac caa agc gaa gta tcc aga ttg ttt gaa aga ttt cat ttg gag 240Ser Asn Gln Ser Glu Val Ser Arg Leu Phe Glu Arg Phe His Leu Glu 65 70 75 80ggc gag ctc cag agg acg act gga aat tca atc cat caa gca caa gac 288Gly Glu Leu Gln Arg Thr Thr Gly Asn Ser Ile His Gln Ala Gln Asp 85 90 95ggt aca acc act aca gct cct tat ggt gac tcc ttg ctg agc gag gag 336Gly Thr Thr Thr Thr Ala Pro Tyr Gly Asp Ser Leu Leu Ser Glu Glu 100 105 110gtt gca agt gca ctt gcg gaa ctc ctc ccc gta tgg tct cag ctg atc 384Val Ala Ser Ala Leu Ala Glu Leu Leu Pro Val Trp Ser Gln Leu Ile 115 120 125gaa gag cat agc ctt caa gac ctc aag gcg agc cct cag gcg aag cgg 432Glu Glu His Ser Leu Gln Asp Leu Lys Ala Ser Pro Gln Ala Lys Arg 130 135 140ctc gac agt gtg agc ttc gcg cac tac tgt gag aag gaa cta aac ttg 480Leu Asp Ser Val Ser Phe Ala His Tyr Cys Glu Lys Glu Leu Asn Leu145 150 155 160cct gct gtt ctc ggc gta gca aac cag atc aca cgc gct ctg ctc ggt 528Pro Ala Val Leu Gly Val Ala Asn Gln Ile Thr Arg Ala Leu Leu Gly 165 170 175gtg gaa gcc cac gag atc agc atg ctt ttt ctc acc gac tac atc aag 576Val Glu Ala His Glu Ile Ser Met Leu Phe Leu Thr Asp Tyr Ile Lys 180 185 190agt gcc acc ggt ctc agt aat att ttc tcg gac aag aaa gac ggc ggg 624Ser Ala Thr Gly Leu Ser Asn Ile Phe Ser Asp Lys Lys Asp Gly Gly 195 200 205cag tat atg cga tgc aaa aca ggt atg cag tcg att tgc cat gcc atg 672Gln Tyr Met Arg Cys Lys Thr Gly Met Gln Ser Ile Cys His Ala Met 210 215 220tca aag gaa ctt gtt cca ggc tca gtg cac ctc aac acc ccc gtc gct 720Ser Lys Glu Leu Val Pro Gly Ser Val His Leu Asn Thr Pro Val Ala225 230 235 240gaa att gag cag tcg gca tcc ggc tgt aca gta cga tcg gcc tcg ggc 768Glu Ile Glu Gln Ser Ala Ser Gly Cys Thr Val Arg Ser Ala Ser Gly 245 250 255gcc gtg ttc cga agc aaa aag gtg gtg gtt tcg tta ccg aca acc ttg 816Ala Val Phe Arg Ser Lys Lys Val Val Val Ser Leu Pro Thr Thr Leu 260 265 270tat ccc acc ttg aca ttt tca cca cct ctt ccc gcc gag aag caa gca 864Tyr Pro Thr Leu Thr Phe Ser Pro Pro Leu Pro Ala Glu Lys Gln Ala 275 280 285ttg gcg gaa aat tct atc ctg ggc tac tat agc aag ata gtc ttc gta 912Leu Ala Glu Asn Ser Ile Leu Gly Tyr Tyr Ser Lys Ile Val Phe Val 290 295 300tgg gac aag ccg tgg tgg cgc gaa caa ggc ttc tcg ggc gtc ctc caa 960Trp Asp Lys Pro Trp Trp Arg Glu Gln Gly Phe Ser Gly Val Leu Gln305 310 315 320tcg agc tgt gac ccc atc tca ttt gcc aga gat acc agc atc gac gtc 1008Ser Ser Cys Asp Pro Ile Ser Phe Ala Arg Asp Thr Ser Ile Asp Val 325 330 335gat cga caa tgg tcc att acc tgt ttc atg gtc gga gac ccg gga cgg 1056Asp Arg Gln Trp Ser Ile Thr Cys Phe Met Val Gly Asp Pro Gly Arg 340 345 350aag tgg tcc caa cag tcc aag cag gta cga caa aag tct gtc tgg gac 1104Lys Trp Ser Gln Gln Ser Lys Gln Val Arg Gln Lys Ser Val Trp Asp 355 360 365caa ctc cgc gca gcc tac gag aac gcc ggg gcc caa gtc cca gag ccg 1152Gln Leu Arg Ala Ala Tyr Glu Asn Ala Gly Ala Gln Val Pro Glu Pro 370 375 380gcc aac gtg ctc gaa atc gag tgg tcg aag cag cag tat ttc caa gga 1200Ala Asn Val Leu Glu Ile Glu Trp Ser Lys Gln Gln Tyr Phe Gln Gly385 390 395 400gct ccg agc gcc gtc tat ggg ctg aac gat ctc atc aca ctg ggt tcg 1248Ala Pro Ser Ala Val Tyr Gly Leu Asn Asp Leu Ile Thr Leu Gly Ser 405 410 415gcg ctc aga acg ccg ttc aag agt gtt cat ttc gtt gga acg gag acg 1296Ala Leu Arg Thr Pro Phe Lys Ser Val His Phe Val Gly Thr Glu Thr 420 425 430tct tta gtt tgg aaa ggg tat atg gaa ggg gcc ata cga tcg ggt caa 1344Ser Leu Val Trp Lys Gly Tyr Met Glu Gly Ala Ile Arg Ser Gly Gln 435 440 445cga ggt gct gca gaa gtt gtg gct agc ctg gtg cca gca gca tag 1389Arg Gly Ala Ala Glu Val Val Ala Ser Leu Val Pro Ala Ala * 450 455 460 17 462 PRT Exophiala spinifera 17Asp Asn Val Ala Asp Val Val Val Val Gly Ala Gly Leu Ser Gly Leu 1 5 10 15Glu Thr Ala Arg Lys Val Gln Ala Ala Gly Leu Ser Cys Leu Val Leu 20 25 30Glu Ala Met Asp Arg Val Gly Gly Lys Thr Leu Ser Val Gln Ser Gly 35 40 45Pro Gly Arg Thr Thr Ile Asn Asp Leu Gly Ala Ala Trp Ile Asn Asp 50 55 60Ser Asn Gln Ser Glu Val Ser Arg Leu Phe Glu Arg Phe His Leu Glu65 70 75 80Gly Glu Leu Gln Arg Thr Thr Gly Asn Ser Ile His Gln Ala Gln Asp 85 90 95Gly Thr Thr Thr Thr Ala Pro Tyr Gly Asp Ser Leu Leu Ser Glu Glu 100 105 110Val Ala Ser Ala Leu Ala Glu Leu Leu Pro Val Trp Ser Gln Leu Ile 115 120 125Glu Glu His Ser Leu Gln Asp Leu Lys Ala Ser Pro Gln Ala Lys Arg 130 135 140Leu Asp Ser Val Ser Phe Ala His Tyr Cys Glu Lys Glu Leu Asn Leu145 150 155 160Pro Ala Val Leu Gly Val Ala Asn Gln Ile Thr Arg Ala Leu Leu Gly 165 170 175Val Glu Ala His Glu Ile Ser Met Leu Phe Leu Thr Asp Tyr Ile Lys 180 185 190Ser Ala Thr Gly Leu Ser Asn Ile Phe Ser Asp Lys Lys Asp Gly Gly 195 200 205Gln Tyr Met Arg Cys Lys Thr Gly Met Gln Ser Ile Cys His Ala Met 210 215 220Ser Lys Glu Leu Val Pro Gly Ser Val His Leu Asn Thr Pro Val Ala225 230 235 240Glu Ile Glu Gln Ser Ala Ser Gly Cys Thr Val Arg Ser Ala Ser Gly 245 250 255Ala Val Phe Arg Ser Lys Lys Val Val Val Ser Leu Pro Thr Thr Leu 260 265 270Tyr Pro Thr Leu Thr Phe Ser Pro Pro Leu Pro Ala Glu Lys Gln Ala 275 280 285Leu Ala Glu Asn Ser Ile Leu Gly Tyr Tyr Ser Lys Ile Val Phe Val 290 295 300Trp Asp Lys Pro Trp Trp Arg Glu Gln Gly Phe Ser Gly Val Leu Gln305 310 315 320Ser Ser Cys Asp Pro Ile Ser Phe Ala Arg Asp Thr Ser Ile Asp Val 325 330 335Asp Arg Gln Trp Ser Ile Thr Cys Phe Met Val Gly Asp Pro Gly Arg 340 345 350Lys Trp Ser Gln Gln Ser Lys Gln Val Arg Gln Lys Ser Val Trp Asp 355 360 365Gln Leu Arg Ala Ala Tyr Glu Asn Ala Gly Ala Gln Val Pro Glu Pro 370 375 380Ala Asn Val Leu Glu Ile Glu Trp Ser Lys Gln Gln Tyr Phe Gln Gly385 390 395 400Ala Pro Ser Ala Val Tyr Gly Leu Asn Asp Leu Ile Thr Leu Gly Ser 405 410 415Ala Leu Arg Thr Pro Phe Lys Ser Val His Phe Val Gly Thr Glu Thr 420 425 430Ser Leu Val Trp Lys Gly Tyr Met Glu Gly Ala Ile Arg Ser Gly Gln 435 440 445Arg Gly Ala Ala Glu Val Val Ala Ser Leu Val Pro Ala Ala 450 455 460 18 1392 DNA Exophiala spinifera CDS (1)...(1392) misc_feature (0)...(0) truncated APAO with additional Lysine 18aaa gac aac gtt gcg gac gtg gta gtg gtg ggc gct ggc ttg agc ggt 48Lys Asp Asn Val Ala Asp Val Val Val Val Gly Ala Gly Leu Ser Gly 1 5 10 15ttg gag acg gca cgc aaa gtc cag gcc gcc ggt ctg tcc tgc ctc gtt 96Leu Glu Thr Ala Arg Lys Val Gln Ala Ala Gly Leu Ser Cys Leu Val 20 25 30ctt gag gcg atg gat cgt gta ggg gga aag act ctg agc gta caa tcg 144Leu Glu Ala Met Asp Arg Val Gly Gly Lys Thr Leu Ser Val Gln Ser 35 40 45ggt ccc ggc agg acg act atc aac gac ctc ggc gct gcg tgg atc aat 192Gly Pro Gly Arg Thr Thr Ile Asn Asp Leu Gly Ala Ala Trp Ile Asn 50 55 60gac agc aac caa agc gaa gta tcc aga ttg ttt gaa aga ttt cat ttg 240Asp Ser Asn Gln Ser Glu Val Ser Arg Leu Phe Glu Arg Phe His Leu 65 70 75 80gag ggc gag ctc cag agg acg act gga aat tca atc cat caa gca caa 288Glu Gly Glu Leu Gln Arg Thr Thr Gly Asn Ser Ile His Gln Ala Gln 85 90 95gac ggt aca acc act aca gct cct tat ggt gac tcc ttg ctg agc gag 336Asp Gly Thr Thr Thr Thr Ala Pro Tyr Gly Asp Ser Leu Leu Ser Glu 100 105 110gag gtt gca agt gca ctt gcg gaa ctc ctc ccc gta tgg tct cag ctg 384Glu Val Ala Ser Ala Leu Ala Glu Leu Leu Pro Val Trp Ser Gln Leu 115 120 125atc gaa gag cat agc ctt caa gac ctc aag gcg agc cct cag gcg aag 432Ile Glu Glu His Ser Leu Gln Asp Leu Lys Ala Ser Pro Gln Ala Lys 130 135 140cgg ctc gac agt gtg agc ttc gcg cac tac tgt gag aag gaa cta aac 480Arg Leu Asp Ser Val Ser Phe Ala His Tyr Cys Glu Lys Glu Leu Asn145 150 155 160ttg cct gct gtt ctc ggc gta gca aac cag atc aca cgc gct ctg ctc 528Leu Pro Ala Val Leu Gly Val Ala Asn Gln Ile Thr Arg Ala Leu Leu 165 170 175ggt gtg gaa gcc cac gag atc agc atg ctt ttt ctc acc gac tac atc 576Gly Val Glu Ala His Glu Ile Ser Met Leu Phe Leu Thr Asp Tyr Ile 180 185 190aag agt gcc acc ggt ctc agt aat att ttc tcg gac aag aaa gac ggc 624Lys Ser Ala Thr Gly Leu Ser Asn Ile Phe Ser Asp Lys Lys Asp Gly 195 200 205ggg cag tat atg cga tgc aaa aca ggt atg cag tcg att tgc cat gcc 672Gly Gln Tyr Met Arg Cys Lys Thr Gly Met Gln Ser Ile Cys His Ala 210 215 220atg tca aag gaa ctt gtt cca ggc tca gtg cac ctc aac acc ccc gtc 720Met Ser Lys Glu Leu Val Pro Gly Ser Val His Leu Asn Thr Pro Val225 230 235 240gct gaa att gag cag tcg gca tcc ggc tgt aca gta cga tcg gcc tcg 768Ala Glu Ile Glu Gln Ser Ala Ser Gly Cys Thr Val Arg Ser Ala Ser 245 250 255ggc gcc gtg ttc cga agc aaa aag gtg gtg gtt tcg tta ccg aca acc 816Gly Ala Val Phe Arg Ser Lys Lys Val Val Val Ser Leu Pro Thr Thr 260 265 270ttg tat ccc acc ttg aca ttt tca cca cct ctt ccc gcc gag aag caa 864Leu Tyr Pro Thr Leu Thr Phe Ser Pro Pro Leu Pro Ala Glu Lys Gln 275 280 285gca ttg gcg gaa aat tct atc ctg ggc tac tat agc aag ata gtc ttc 912Ala Leu Ala Glu Asn Ser Ile Leu Gly Tyr Tyr Ser Lys Ile Val Phe 290 295 300gta tgg gac aag ccg tgg tgg cgc gaa caa ggc ttc tcg ggc gtc ctc 960Val Trp Asp Lys Pro Trp Trp Arg Glu Gln Gly Phe Ser Gly Val Leu305 310 315 320caa tcg agc tgt gac ccc atc tca ttt gcc aga gat acc agc atc gac 1008Gln Ser Ser Cys Asp Pro Ile Ser Phe Ala Arg Asp Thr Ser Ile Asp 325 330 335gtc gat cga caa tgg tcc att acc tgt ttc atg gtc gga gac ccg gga 1056Val Asp Arg Gln Trp Ser Ile Thr Cys Phe Met Val Gly Asp Pro Gly 340 345 350cgg aag tgg tcc caa cag tcc aag cag gta cga caa aag tct gtc tgg 1104Arg Lys Trp Ser Gln Gln Ser Lys Gln Val Arg Gln Lys Ser Val Trp 355 360 365gac caa ctc cgc gca gcc tac gag aac gcc ggg gcc caa gtc cca gag 1152Asp Gln Leu Arg Ala Ala Tyr Glu Asn Ala Gly Ala Gln Val Pro Glu 370 375 380ccg gcc aac gtg ctc gaa atc gag tgg tcg aag cag cag tat ttc caa 1200Pro Ala Asn Val Leu Glu Ile Glu Trp Ser Lys Gln Gln Tyr Phe Gln385 390 395 400gga gct ccg agc gcc gtc tat ggg ctg aac gat ctc atc aca ctg ggt 1248Gly Ala Pro Ser Ala Val Tyr Gly Leu Asn Asp Leu Ile Thr Leu Gly 405 410 415tcg gcg ctc aga acg ccg ttc aag agt gtt cat ttc gtt gga acg gag 1296Ser Ala Leu Arg Thr Pro Phe Lys Ser Val His Phe Val Gly Thr Glu 420 425 430acg tct tta gtt tgg aaa ggg tat atg gaa ggg gcc ata cga tcg ggt 1344Thr Ser Leu Val Trp Lys Gly Tyr Met Glu Gly Ala Ile Arg Ser Gly 435 440 445caa cga ggt gct gca gaa gtt gtg gct agc ctg gtg cca gca gca tag 1392Gln Arg Gly Ala Ala Glu Val Val Ala Ser Leu Val Pro Ala Ala * 450 455 460 19 463 PRT Exophiala spinifera 19Lys Asp Asn Val Ala Asp Val Val Val Val Gly Ala Gly Leu Ser Gly 1 5 10 15Leu Glu Thr Ala Arg Lys Val Gln Ala Ala Gly Leu Ser Cys Leu Val 20 25 30Leu Glu Ala Met Asp Arg Val Gly Gly Lys Thr Leu Ser Val Gln Ser 35 40 45Gly Pro Gly Arg Thr Thr Ile Asn Asp Leu Gly Ala Ala Trp Ile Asn 50 55 60Asp Ser Asn Gln Ser Glu Val Ser Arg Leu Phe Glu Arg Phe His Leu65 70 75 80Glu Gly Glu Leu Gln Arg Thr Thr Gly Asn Ser Ile His Gln Ala Gln 85 90 95Asp Gly Thr Thr Thr Thr Ala Pro Tyr Gly Asp Ser Leu Leu Ser Glu 100 105 110Glu Val Ala Ser Ala Leu Ala Glu Leu Leu Pro Val Trp Ser Gln Leu 115 120 125Ile Glu Glu His Ser Leu Gln Asp Leu Lys Ala Ser Pro Gln Ala Lys 130 135 140Arg Leu Asp Ser Val Ser Phe Ala His Tyr Cys Glu Lys Glu Leu Asn145 150 155 160Leu Pro Ala Val Leu Gly Val Ala Asn Gln Ile Thr Arg Ala Leu Leu 165 170 175Gly Val Glu Ala His Glu Ile Ser Met Leu Phe Leu Thr Asp Tyr Ile 180 185 190Lys Ser Ala Thr Gly Leu Ser Asn Ile Phe Ser Asp Lys Lys Asp Gly 195 200 205Gly Gln Tyr Met Arg Cys Lys Thr Gly Met Gln Ser Ile Cys His Ala 210 215 220Met Ser Lys Glu Leu Val Pro Gly Ser Val His Leu Asn Thr Pro Val225 230 235 240Ala Glu Ile Glu Gln Ser Ala Ser Gly Cys Thr Val Arg Ser Ala Ser 245 250 255Gly Ala Val Phe Arg Ser Lys Lys Val Val Val Ser Leu Pro Thr Thr 260 265 270Leu Tyr Pro Thr Leu Thr Phe Ser Pro Pro Leu Pro Ala Glu Lys Gln 275 280 285Ala Leu Ala Glu Asn Ser Ile Leu Gly Tyr Tyr Ser Lys Ile Val Phe 290 295 300Val Trp Asp Lys Pro Trp Trp Arg Glu Gln Gly Phe Ser Gly Val Leu305 310 315 320Gln Ser Ser Cys Asp Pro Ile Ser Phe Ala Arg Asp Thr Ser Ile Asp 325 330 335Val Asp Arg Gln Trp Ser Ile Thr Cys Phe Met Val Gly Asp Pro Gly 340 345 350Arg Lys Trp Ser Gln Gln Ser Lys Gln Val Arg Gln Lys Ser Val Trp 355 360 365Asp Gln Leu Arg Ala Ala Tyr Glu Asn Ala Gly Ala Gln Val Pro Glu 370 375 380Pro Ala Asn Val Leu Glu Ile Glu Trp Ser Lys Gln Gln Tyr Phe Gln385 390 395 400Gly Ala Pro Ser Ala Val Tyr Gly Leu Asn Asp Leu Ile Thr Leu Gly 405 410 415Ser Ala Leu Arg Thr Pro Phe Lys Ser Val His Phe Val Gly Thr Glu 420 425 430Thr Ser Leu Val Trp Lys Gly Tyr Met Glu Gly Ala Ile Arg Ser Gly 435 440 445Gln Arg Gly Ala Ala Glu Val Val Ala Ser Leu Val Pro Ala Ala 450 455 460 20 1803 DNA Exophiala spinifera CDS (1)...(1800) misc_feature (0)...(0) full-length APAO 20atg gca ctt gca ccg agc tac atc aat ccc cca aac gtc gcc tcc cca 48Met Ala Leu Ala Pro Ser Tyr Ile Asn Pro Pro Asn Val Ala Ser Pro 1 5 10 15gca ggg tat tct cac gtc ggc gta ggc cca gac gga ggg agg tat gtg 96Ala Gly Tyr Ser His Val Gly Val Gly Pro Asp Gly Gly Arg Tyr Val 20 25 30aca ata gct gga cag att gga caa gac gct tcg ggc gtg aca gac cct 144Thr Ile Ala Gly Gln Ile Gly Gln Asp Ala Ser Gly Val Thr Asp Pro 35 40 45gcc tac gag aaa cag gtt gcc caa gca ttc gcc aat ctg cga gct tgc 192Ala Tyr Glu Lys Gln Val Ala Gln Ala Phe Ala Asn Leu Arg Ala Cys 50 55 60ctt gct gca gtt gga gcc act tca aac gac gtc acc aag ctc aat tac 240Leu Ala Ala Val Gly Ala Thr Ser Asn Asp Val Thr Lys Leu Asn Tyr 65 70 75 80tac atc gtc gac tac gcc ccg agc aaa ctc acc gca att gga gat ggg 288Tyr Ile Val Asp Tyr Ala Pro Ser Lys Leu Thr Ala Ile Gly Asp Gly 85 90 95ctg aag gct acc ttt gcc ctt gac agg ctc cct cct tgc acg ctg gtg 336Leu Lys Ala Thr Phe Ala Leu Asp Arg Leu Pro Pro Cys Thr Leu Val 100 105 110cca gtg tcg gcc ttg tct tca cct gaa tac ctc ttt gag gtt gat gcc 384Pro Val Ser Ala Leu Ser Ser Pro Glu Tyr Leu Phe Glu Val Asp Ala 115 120 125acg gcg ctg gtg ccg gga cac acg acc cca gac aac gtt gcg gac gtg 432Thr Ala Leu Val Pro Gly His Thr Thr Pro Asp Asn Val Ala Asp Val 130 135 140gta gtg gtg ggc gct ggc ttg agc ggt ttg gag acg gca cgc aaa gtc 480Val Val Val Gly Ala Gly Leu Ser Gly Leu Glu Thr Ala Arg Lys Val145 150 155 160cag gcc gcc ggt ctg tcc tgc ctc gtt ctt gag gcg atg gat cgt gta 528Gln Ala Ala Gly Leu Ser Cys Leu Val Leu Glu Ala Met Asp Arg Val 165 170 175ggg gga aag act ctg agc gta caa tcg ggt ccc ggc agg acg act atc 576Gly Gly Lys Thr Leu Ser Val Gln Ser Gly Pro Gly Arg Thr Thr Ile 180 185 190aac gac ctc ggc gct gcg tgg atc aat gac agc aac caa agc gaa gta 624Asn Asp Leu Gly Ala Ala Trp Ile Asn Asp Ser Asn Gln Ser Glu Val 195 200 205tcc aga ttg ttt gaa aga ttt cat ttg gag ggc gag ctc cag agg acg 672Ser Arg Leu Phe Glu Arg Phe His Leu Glu Gly Glu Leu Gln Arg Thr 210 215 220act gga aat tca atc cat caa gca caa gac ggt aca acc act aca gct 720Thr Gly Asn Ser Ile His Gln Ala Gln Asp Gly Thr Thr Thr Thr Ala225 230 235 240cct tat ggt gac tcc ttg ctg agc gag gag gtt gca agt gca ctt gcg 768Pro Tyr Gly Asp Ser Leu Leu Ser Glu Glu Val Ala Ser Ala Leu Ala 245 250 255gaa ctc ctc ccc gta tgg tct cag ctg atc gaa gag cat agc ctt caa 816Glu Leu Leu Pro Val Trp Ser Gln Leu Ile Glu Glu His Ser Leu Gln 260 265 270gac ctc aag gcg agc cct cag gcg aag cgg ctc gac agt gtg agc ttc 864Asp Leu Lys Ala Ser Pro Gln Ala Lys Arg Leu Asp Ser Val Ser Phe 275 280 285gcg cac tac tgt gag aag gaa cta aac ttg cct gct gtt ctc ggc gta 912Ala His Tyr Cys Glu Lys Glu Leu Asn Leu Pro Ala Val Leu Gly Val 290 295 300gca aac cag atc aca cgc gct ctg ctc ggt gtg gaa gcc cac gag atc 960Ala Asn Gln Ile Thr Arg Ala Leu Leu Gly Val Glu Ala His Glu Ile305 310 315 320agc atg ctt ttt ctc acc gac tac atc aag agt gcc acc ggt ctc agt 1008Ser Met Leu Phe Leu Thr Asp Tyr Ile Lys Ser Ala Thr Gly Leu Ser 325 330 335aat att ttc tcg gac aag aaa gac ggc ggg cag tat atg cga tgc aaa 1056Asn Ile Phe Ser Asp Lys Lys Asp Gly Gly Gln Tyr Met Arg Cys Lys 340 345 350aca ggt atg cag tcg att tgc cat gcc atg tca aag gaa ctt gtt cca 1104Thr Gly Met Gln Ser Ile Cys His Ala Met Ser Lys Glu Leu Val Pro 355 360 365ggc tca gtg cac ctc aac acc ccc gtc gct gaa att gag cag tcg gca 1152Gly Ser Val His Leu Asn Thr Pro Val Ala Glu Ile Glu Gln Ser Ala 370 375 380tcc ggc tgt aca gta cga tcg gcc tcg ggc gcc gtg ttc cga agc aaa 1200Ser Gly Cys Thr Val Arg Ser Ala Ser Gly Ala Val Phe Arg Ser Lys385 390 395 400aag gtg gtg gtt tcg tta ccg aca acc ttg tat ccc acc ttg aca ttt 1248Lys Val Val Val Ser Leu Pro Thr Thr Leu Tyr Pro Thr Leu Thr Phe 405 410 415tca cca cct ctt ccc gcc gag aag caa gca ttg gcg gaa aat tct atc 1296Ser Pro Pro Leu Pro Ala Glu Lys Gln Ala Leu Ala Glu Asn Ser Ile 420 425 430ctg ggc tac tat agc aag ata gtc ttc gta tgg gac aag ccg tgg tgg 1344Leu Gly Tyr Tyr Ser Lys Ile Val Phe Val Trp Asp Lys Pro Trp Trp 435 440 445cgc gaa caa ggc ttc tcg ggc gtc ctc caa tcg agc tgt gac ccc atc 1392Arg Glu Gln Gly Phe Ser Gly Val Leu Gln Ser Ser Cys Asp Pro Ile 450 455 460tca ttt gcc aga gat acc agc atc gac gtc gat cga caa tgg tcc att 1440Ser Phe Ala Arg Asp Thr Ser Ile Asp Val Asp Arg Gln Trp Ser Ile465 470 475 480acc tgt ttc atg gtc gga gac ccg gga cgg aag tgg tcc caa cag tcc 1488Thr Cys Phe Met Val Gly Asp Pro Gly Arg Lys Trp Ser Gln Gln Ser 485 490 495aag cag gta cga caa aag tct gtc tgg gac caa ctc cgc gca gcc tac 1536Lys Gln Val Arg Gln Lys Ser Val Trp Asp Gln Leu Arg Ala Ala Tyr 500 505 510gag aac gcc ggg gcc caa gtc cca gag ccg gcc aac gtg ctc gaa atc 1584Glu Asn Ala Gly Ala Gln Val Pro Glu Pro Ala Asn Val Leu Glu Ile 515 520 525gag tgg tcg aag cag cag tat ttc caa gga gct ccg agc gcc gtc tat 1632Glu Trp Ser Lys Gln Gln Tyr Phe Gln Gly Ala Pro Ser Ala Val Tyr 530 535 540ggg ctg aac gat ctc atc aca ctg ggt tcg gcg ctc aga acg ccg ttc 1680Gly Leu Asn Asp Leu Ile Thr Leu Gly Ser Ala Leu Arg Thr Pro Phe545 550 555 560aag agt gtt cat ttc gtt gga acg gag acg tct tta gtt tgg aaa ggg 1728Lys Ser Val His Phe Val Gly Thr Glu Thr Ser Leu Val Trp Lys Gly 565 570 575tat atg gaa ggg gcc ata cga tcg ggt caa cga ggt gct gca gaa gtt 1776Tyr Met Glu Gly Ala Ile Arg Ser Gly Gln Arg Gly Ala Ala Glu Val 580 585 590gtg gct agc ctg gtg cca gca gca tag 1803Val Ala Ser Leu Val Pro Ala Ala 595 600 21 600 PRT Exophiala spinifera 21Met Ala Leu Ala Pro Ser Tyr Ile Asn Pro Pro Asn Val Ala Ser Pro 1 5 10 15Ala Gly Tyr Ser His Val Gly Val Gly Pro Asp Gly Gly Arg Tyr Val 20 25 30Thr Ile Ala Gly Gln Ile Gly Gln Asp Ala Ser Gly Val Thr Asp Pro 35 40 45Ala Tyr Glu Lys Gln Val Ala Gln Ala Phe Ala Asn Leu Arg Ala Cys 50 55 60Leu Ala Ala Val Gly Ala Thr Ser Asn Asp Val Thr Lys Leu Asn Tyr65 70 75 80Tyr Ile Val Asp Tyr Ala Pro Ser Lys Leu Thr Ala Ile Gly Asp Gly 85 90 95Leu Lys Ala Thr Phe Ala Leu Asp Arg Leu Pro Pro Cys Thr Leu Val 100 105 110Pro Val Ser Ala Leu Ser Ser Pro Glu Tyr Leu Phe Glu Val Asp Ala 115 120 125Thr Ala Leu Val Pro Gly His Thr Thr Pro Asp Asn Val Ala Asp Val 130 135 140Val Val Val Gly Ala Gly Leu Ser Gly Leu Glu Thr Ala Arg Lys Val145 150 155 160Gln Ala Ala Gly Leu Ser Cys Leu Val Leu Glu Ala Met Asp Arg Val 165 170 175Gly Gly Lys Thr Leu Ser Val Gln Ser Gly Pro Gly Arg Thr Thr Ile 180 185 190Asn Asp Leu Gly Ala Ala Trp Ile Asn Asp Ser Asn Gln Ser Glu Val 195 200 205Ser Arg Leu Phe Glu Arg Phe His Leu Glu Gly Glu Leu Gln Arg Thr 210 215 220Thr Gly Asn Ser Ile His Gln Ala Gln Asp Gly Thr Thr Thr Thr Ala225 230 235 240Pro Tyr Gly Asp Ser Leu Leu Ser Glu Glu Val Ala Ser Ala Leu Ala 245 250 255Glu Leu Leu Pro Val Trp Ser Gln Leu Ile Glu Glu His Ser Leu Gln 260 265 270Asp Leu Lys Ala Ser Pro Gln Ala Lys Arg Leu Asp Ser Val Ser Phe 275 280 285Ala His Tyr Cys Glu Lys Glu Leu Asn Leu Pro Ala Val Leu Gly Val 290 295 300Ala Asn Gln Ile Thr Arg Ala Leu Leu Gly Val Glu Ala His Glu Ile305 310 315 320Ser Met Leu Phe Leu Thr Asp Tyr Ile Lys Ser Ala Thr Gly Leu Ser 325 330 335Asn Ile Phe Ser Asp Lys Lys Asp Gly Gly Gln Tyr Met Arg Cys Lys 340 345 350Thr Gly Met Gln Ser Ile Cys His Ala Met Ser Lys Glu Leu Val Pro 355 360 365Gly Ser Val His Leu Asn Thr Pro Val Ala Glu Ile Glu Gln Ser Ala 370 375 380Ser Gly Cys Thr Val Arg Ser Ala Ser Gly Ala Val Phe Arg Ser Lys385 390 395 400Lys Val Val Val Ser Leu Pro Thr Thr Leu Tyr Pro Thr Leu Thr Phe 405 410 415Ser Pro Pro Leu Pro Ala Glu Lys Gln Ala Leu Ala Glu Asn Ser Ile 420 425 430Leu Gly Tyr Tyr Ser Lys Ile Val Phe Val Trp Asp Lys Pro Trp Trp 435 440 445Arg Glu Gln Gly Phe Ser Gly Val Leu Gln Ser Ser Cys Asp Pro Ile 450 455 460Ser Phe Ala Arg Asp Thr Ser Ile Asp Val Asp Arg Gln Trp Ser Ile465 470 475 480Thr Cys Phe Met Val Gly Asp Pro Gly Arg Lys Trp Ser Gln Gln Ser 485 490 495Lys Gln Val Arg Gln Lys Ser Val Trp Asp Gln Leu Arg Ala Ala Tyr 500 505 510Glu Asn Ala Gly Ala Gln Val Pro Glu Pro Ala Asn Val Leu Glu Ile 515 520 525Glu Trp Ser Lys Gln Gln Tyr Phe Gln Gly Ala Pro Ser Ala Val Tyr 530 535 540Gly Leu Asn Asp Leu Ile Thr Leu Gly Ser Ala Leu Arg Thr Pro Phe545 550 555 560Lys Ser Val His Phe Val Gly Thr Glu Thr Ser Leu Val Trp Lys Gly 565 570 575Tyr Met Glu Gly Ala Ile Arg Ser Gly Gln Arg Gly Ala Ala Glu Val 580 585 590Val Ala Ser Leu Val Pro Ala Ala 595 600 22 1803 DNA Exophiala spinifera CDS (1)...(1803) misc_feature (0)...(0) isolate ESP002_C2 22atg gca ctt gca ccg agc tac atc aat ccc cca aac gtc gcc tcc cca 48Met Ala Leu Ala Pro Ser Tyr Ile Asn Pro Pro Asn Val Ala Ser Pro 1 5 10 15gca ggg tat tcc cac atc ggc gta ggc cca aac gaa gcg agg tat gtg 96Ala Gly Tyr Ser His Ile Gly Val Gly Pro Asn Glu Ala Arg Tyr Val 20 25 30aca ata gct gga cag att gga caa gac gct ttg ggc gtg aca gac cca 144Thr Ile Ala Gly Gln Ile Gly Gln Asp Ala Leu Gly Val Thr Asp Pro 35 40 45gcc tac gag aaa cag gtt gcc caa gca ttc gcc aat ctg cga gct tgc 192Ala Tyr Glu Lys Gln Val Ala Gln Ala Phe Ala Asn Leu Arg Ala Cys 50 55 60ctt gct gca gtt gga gcc tct tca aac gac gtc acc aag ctc aat tac 240Leu Ala Ala Val Gly Ala Ser Ser Asn Asp Val Thr Lys Leu Asn Tyr 65 70 75 80tac atc gtc gac tac gcc ccg agc aaa ctc acc gca att gga gat ggg 288Tyr Ile Val Asp Tyr Ala Pro Ser Lys Leu Thr Ala Ile Gly Asp Gly 85 90 95ctg aag tct acc ttt gcc ctt gac agg ctc cct cct tgc acg ctg gtg 336Leu Lys Ser Thr Phe Ala Leu Asp Arg Leu Pro Pro Cys Thr Leu Val 100 105 110cca gta ccg gcc ttg gct tca cct gaa tac ctc ttt gag gtt gat gcc 384Pro Val Pro Ala Leu Ala Ser Pro Glu Tyr Leu Phe Glu Val Asp Ala 115 120 125acg gcg ctg gtg cca gga cac tcg acc cca gac aac gtt gcg gac gtg 432Thr Ala Leu Val Pro Gly His Ser Thr Pro Asp Asn Val Ala Asp Val 130 135 140gta gtg gtg ggc gct ggc ttg agc ggt ttg gag acg gca cgc aaa gtc 480Val Val Val Gly Ala Gly Leu Ser Gly Leu Glu Thr Ala Arg Lys Val145 150 155 160cag gcc gcc ggt ctg tcc tgc ctc gtt ctt gag gcg atg gat cgt gta 528Gln Ala Ala Gly Leu Ser Cys Leu Val Leu Glu Ala Met Asp Arg Val 165 170 175ggg gga aag act ctg agc gta caa tcg ggt ccc ggc agg acg act atc 576Gly Gly Lys Thr Leu Ser Val Gln Ser Gly Pro Gly Arg Thr Thr Ile 180 185 190aac gac ctc ggc gct gcg tgg atc aat gac agc aac caa agc gaa gta 624Asn Asp Leu Gly Ala Ala Trp Ile Asn Asp Ser Asn Gln Ser Glu Val 195 200 205tcc aga ttg ttt gaa aga ttt cat ttg gag ggc gag ctc cag agg acg 672Ser Arg Leu Phe Glu Arg Phe His Leu Glu Gly Glu Leu Gln Arg Thr 210 215 220acc gga aat tca atc cat caa gca caa gac ggt aca acc act aca gct 720Thr Gly Asn Ser Ile His Gln Ala Gln Asp Gly Thr Thr Thr Thr Ala225 230 235 240cct tat ggt gac tcc ccg ctg agc gag gag gtt gca agt gca ctt gcg 768Pro Tyr Gly Asp Ser Pro Leu Ser Glu Glu Val Ala Ser Ala Leu Ala 245 250 255gaa ctc ctc ccc gta tgg tct cag ctg atc gaa gag tat agc ctt gaa 816Glu Leu Leu Pro Val Trp Ser Gln Leu Ile Glu Glu Tyr Ser Leu Glu 260 265 270gac ccc aag gcg agc cct cag gcg aag cgg ctc gac agt gtg agc ttc 864Asp Pro Lys Ala Ser Pro Gln Ala Lys Arg Leu Asp Ser Val Ser Phe 275 280 285gcg cac tac tgt gag aag gac cta aac ttg cct gct gtt ctc agc gtg 912Ala His Tyr Cys Glu Lys Asp Leu Asn Leu Pro Ala Val Leu Ser Val 290 295 300gca aac cag atc aca cgc gct ctg ctc ggt gtg gaa gcc cac gag atc 960Ala Asn Gln Ile Thr Arg Ala Leu Leu Gly Val Glu Ala His Glu Ile305 310 315 320agc atg ctt ttt ctc acc gac tac atc aag agt gcc acc ggt ctc agt 1008Ser Met Leu Phe Leu Thr Asp Tyr Ile Lys Ser Ala Thr Gly Leu Ser 325 330 335aat att gtc tcg gac aag aaa gac ggc ggg cag tat atg cga tgc aaa 1056Asn Ile Val Ser Asp Lys Lys Asp Gly Gly Gln Tyr Met Arg Cys Lys 340 345 350aca ggt atg cag tcg att tgc cat gcc atg tca aag gaa ctt gtt cca 1104Thr Gly Met Gln Ser Ile Cys His Ala Met Ser Lys Glu Leu Val Pro 355 360 365ggc tca gtg cac ctc aac acc ccc gtc gct gga att gag cag tcg gcg 1152Gly Ser Val His Leu Asn Thr Pro Val Ala Gly Ile Glu Gln Ser Ala 370 375 380tcc ggc tgt ata gta cga tcg gcc tcg ggc gcc gtg ttc cga agc aaa 1200Ser Gly Cys Ile Val Arg Ser Ala Ser Gly Ala Val Phe Arg Ser Lys385 390 395 400aag gtg gtg gtt tcg tta ccg aca aca ttg tat ccc acc ttg aca ttt 1248Lys Val Val Val Ser Leu Pro Thr Thr Leu Tyr Pro Thr Leu Thr Phe 405 410 415tca cca cct ctt ccc gcc gag aag caa gca ttg gcg gaa aaa tct atc 1296Ser Pro Pro Leu Pro Ala Glu Lys Gln Ala Leu Ala Glu Lys Ser Ile 420 425 430ctc ggc tac tat agc aag ata gtc ttc gta tgg gac aac ccg tgg tgg 1344Leu Gly Tyr Tyr Ser Lys Ile Val Phe Val Trp Asp Asn Pro Trp Trp 435 440 445cgc gaa caa ggc ttc tcg ggc gtc ctc caa tcg agc tgt gac ccc atc 1392Arg Glu Gln Gly Phe Ser Gly Val Leu Gln Ser Ser Cys Asp Pro Ile 450 455 460tca ttt gcc aga gat acc agc atc gaa gtc gat cgg caa tgg tcc att 1440Ser Phe Ala Arg Asp Thr Ser Ile Glu Val Asp Arg Gln Trp Ser Ile465 470 475 480acc tgt ttc atg gtc gga gac ccg gga cgg aag tgg tcc caa cag tcc 1488Thr Cys Phe Met Val Gly Asp Pro Gly Arg Lys Trp Ser Gln Gln Ser 485 490 495aag cag gta cga caa aag tct gtc tgg gac caa ctc cgc gca gcc tac 1536Lys Gln Val Arg Gln Lys Ser Val Trp Asp Gln Leu Arg Ala Ala Tyr 500 505 510gag aac gcc ggg gcc caa gtc cca gag ccg gcc aac gtg ctc gaa atc 1584Glu Asn Ala Gly Ala Gln Val Pro Glu Pro Ala Asn Val Leu Glu Ile 515 520 525gag tgg tcg aag cag cag tat ttc caa gga gct ccg agc gcc gtc tat 1632Glu Trp Ser Lys Gln Gln Tyr Phe Gln Gly Ala Pro Ser Ala Val Tyr 530 535 540ggg ctg aac gat ctc atc aca ctg ggt tcg gcg ctc aga acg ccg ttc 1680Gly Leu Asn Asp Leu Ile Thr Leu Gly Ser Ala Leu Arg Thr Pro Phe545 550 555 560aag tgt gtt cat ttc gtt gga acg gag acg tct tta gtt tgg aaa ggg 1728Lys Cys Val His Phe Val Gly Thr Glu Thr Ser Leu Val Trp Lys Gly 565 570 575tat atg gaa ggg gcc ata cga tcg ggt caa cga ggt gct gca gaa gtt 1776Tyr Met Glu Gly Ala Ile Arg Ser Gly Gln Arg Gly Ala Ala Glu Val 580 585 590gtg gct agc ctg gtg cca gca gca tag 1803Val Ala Ser Leu Val Pro Ala Ala * 595 600 23 600 PRT Exophiala spinifera 23Met Ala Leu Ala Pro Ser Tyr Ile Asn Pro Pro Asn Val Ala Ser Pro 1 5 10 15Ala Gly Tyr Ser His Ile Gly Val Gly Pro Asn Glu Ala Arg Tyr Val 20 25 30Thr Ile Ala Gly Gln Ile Gly Gln Asp Ala Leu Gly Val Thr Asp Pro 35 40 45Ala Tyr Glu Lys Gln Val Ala Gln Ala Phe Ala Asn Leu Arg Ala Cys 50 55 60Leu Ala Ala Val Gly Ala Ser Ser Asn Asp Val Thr Lys Leu Asn Tyr65 70 75 80Tyr Ile Val Asp Tyr Ala Pro Ser Lys Leu Thr Ala Ile Gly Asp Gly 85 90 95Leu Lys Ser Thr Phe Ala Leu Asp Arg Leu Pro Pro Cys Thr Leu Val 100 105 110Pro Val Pro Ala Leu Ala Ser Pro Glu Tyr Leu Phe Glu Val Asp Ala 115 120 125Thr Ala Leu Val Pro Gly His Ser Thr Pro Asp Asn Val Ala Asp Val 130 135 140Val Val Val Gly Ala Gly Leu Ser Gly Leu Glu Thr Ala Arg Lys Val145 150 155 160Gln Ala Ala Gly Leu Ser Cys Leu Val Leu Glu Ala Met Asp Arg Val 165 170 175Gly Gly Lys Thr Leu Ser Val Gln Ser Gly Pro Gly Arg Thr Thr Ile 180 185 190Asn Asp Leu Gly Ala Ala Trp Ile Asn Asp Ser Asn Gln Ser Glu Val 195 200 205Ser Arg Leu Phe Glu Arg Phe His Leu Glu Gly Glu Leu Gln Arg Thr 210 215 220Thr Gly Asn Ser Ile His Gln Ala Gln Asp Gly Thr Thr Thr Thr Ala225 230 235 240Pro Tyr Gly Asp Ser Pro Leu Ser Glu Glu Val Ala Ser Ala Leu Ala 245 250 255Glu Leu Leu Pro Val Trp Ser Gln Leu Ile Glu Glu Tyr Ser Leu Glu 260 265 270Asp Pro Lys Ala Ser Pro Gln Ala Lys Arg Leu Asp Ser Val Ser Phe 275 280 285Ala His Tyr Cys Glu Lys Asp Leu Asn Leu Pro Ala Val Leu Ser Val 290 295 300Ala Asn Gln Ile Thr Arg Ala Leu Leu Gly Val Glu Ala His Glu Ile305 310 315 320Ser Met Leu Phe Leu Thr Asp Tyr Ile Lys Ser Ala Thr Gly Leu Ser 325 330 335Asn Ile Val Ser Asp Lys Lys Asp Gly Gly Gln Tyr Met Arg Cys Lys 340 345 350Thr Gly Met Gln Ser Ile Cys His Ala Met Ser Lys Glu Leu Val Pro 355 360 365Gly Ser Val His Leu Asn Thr Pro Val Ala Gly Ile Glu Gln Ser Ala 370 375 380Ser Gly Cys Ile Val Arg Ser Ala Ser Gly Ala Val Phe Arg Ser Lys385 390 395 400Lys Val Val Val Ser Leu Pro Thr Thr Leu Tyr Pro Thr Leu Thr Phe 405 410 415Ser Pro Pro Leu Pro Ala Glu Lys Gln Ala Leu Ala Glu Lys Ser Ile 420 425 430Leu Gly Tyr Tyr Ser Lys Ile Val Phe Val Trp Asp Asn Pro Trp Trp 435 440 445Arg Glu Gln Gly Phe Ser Gly Val Leu Gln Ser Ser Cys Asp Pro Ile 450 455 460Ser Phe Ala Arg Asp Thr Ser Ile Glu Val Asp Arg Gln Trp Ser Ile465 470 475 480Thr Cys Phe Met Val Gly Asp Pro Gly Arg Lys Trp Ser Gln Gln Ser 485 490 495Lys Gln Val Arg Gln Lys Ser Val Trp Asp Gln Leu Arg Ala Ala Tyr 500 505 510Glu Asn Ala Gly Ala Gln Val Pro Glu Pro Ala Asn Val Leu Glu Ile 515 520 525Glu Trp Ser Lys Gln Gln Tyr Phe Gln Gly Ala Pro Ser Ala Val Tyr 530 535 540Gly Leu Asn Asp Leu Ile Thr Leu Gly Ser Ala Leu Arg Thr Pro Phe545 550 555 560Lys Cys Val His Phe Val Gly Thr Glu Thr Ser Leu Val Trp Lys Gly 565 570 575Tyr Met Glu Gly Ala Ile Arg Ser Gly Gln Arg Gly Ala Ala Glu Val 580 585 590Val Ala Ser Leu Val Pro Ala Ala 595 600 24 1803 DNA Exophiala spinifera CDS (1)...(1803) misc_feature (0)...(0) isolate ESP002_C3 24atg gca ctt gca ccg agc tac atc aat ccc cca aac gtc gcc tcc cca 48Met Ala Leu Ala Pro Ser Tyr Ile Asn Pro Pro Asn Val Ala Ser Pro 1 5 10 15gca ggg tat tcc cac atc ggc gta ggc cca aac gaa gcg agg tat gtg 96Ala Gly Tyr Ser His Ile Gly Val Gly Pro Asn Glu Ala Arg Tyr Val 20 25 30aca ata gct gga cag att gga caa gac gct ttg ggc gtg aca gac cca 144Thr Ile Ala Gly Gln Ile Gly Gln Asp Ala Leu Gly Val Thr Asp Pro 35 40 45gcc tac gag aaa cag gtt gcc caa gca ttc gcc aat ctg cga gct tgc 192Ala Tyr Glu Lys Gln Val Ala Gln Ala Phe Ala Asn Leu Arg Ala Cys 50 55 60ctt gct gca gtt gga gcc tct tca aac gac gtc acc aag ctc aat tac 240Leu Ala Ala Val Gly Ala Ser Ser Asn Asp Val Thr Lys Leu Asn Tyr 65 70 75 80tac atc gtc gac tac gcc ccg agc aaa ctc acc gca att gga gat ggg 288Tyr Ile Val Asp Tyr Ala Pro Ser Lys Leu Thr Ala Ile Gly Asp Gly 85 90 95ctg aag tct acc ttt gcc ctt gac agg ctc cct cct tgc acg ctg gtg 336Leu Lys Ser Thr Phe Ala Leu Asp Arg Leu Pro Pro Cys Thr Leu Val 100 105 110cca gta ccg gcc ttg gct tca cct gaa tac ctc ttt gag gtt gac gcc 384Pro Val Pro Ala Leu Ala Ser Pro Glu Tyr Leu Phe Glu Val Asp Ala 115 120 125acg gcg ctg gtg cca gga cac tcg acc cca gac aac gtt gcg gac gtg 432Thr Ala Leu Val Pro Gly His Ser Thr Pro Asp Asn Val Ala Asp Val 130 135 140gta gtg gtg ggc gct ggc ttg agc ggc ttg gag acg gca cgc aaa gtc 480Val Val Val Gly Ala Gly Leu Ser Gly Leu Glu Thr Ala Arg Lys Val145 150 155 160cag gcc gcc ggt ctg tcc tgc ctc gtt ctt gag gcg atg gat cgt gta 528Gln Ala Ala Gly Leu Ser Cys Leu Val Leu Glu Ala Met Asp Arg Val 165 170 175ggg gga aag act ctg agc gta caa tcg ggt ccc ggc agg acg act atc 576Gly Gly Lys Thr Leu Ser Val Gln Ser Gly Pro Gly Arg Thr Thr Ile 180 185 190aac gac ctc ggc gct gcg tgg atc aat gac agc aac caa agc gaa gta 624Asn Asp Leu Gly Ala Ala Trp Ile Asn Asp Ser Asn Gln Ser Glu Val 195 200 205tcc aga ttg ttt gaa aga ttt cat ttg gag ggc gag ctc cag agg acg 672Ser Arg Leu Phe Glu Arg Phe His Leu Glu Gly Glu Leu Gln Arg Thr 210 215 220acc gga aat tca atc cat caa gca caa gac ggt aca acc act aca gct 720Thr Gly Asn Ser Ile His Gln Ala Gln Asp Gly Thr Thr Thr Thr Ala225 230 235 240cct tat ggt gac tcc ccg ctg agc gag gag gtt gca agt gca ctt gcg 768Pro Tyr Gly Asp Ser Pro Leu Ser Glu Glu Val Ala Ser Ala Leu Ala 245 250 255gaa ctc ctc ccc gta tgg tct cag ctg atc gaa gag tat agc ctt gaa 816Glu Leu Leu Pro Val Trp Ser Gln Leu Ile Glu Glu Tyr Ser Leu Glu 260 265 270gac ccc aag gcg agc cct cag gcg aag cgg ctc gac agt gtg agc ttc 864Asp Pro Lys Ala Ser Pro Gln Ala Lys Arg Leu Asp Ser Val Ser Phe 275 280 285gcg cac tac tgt gag aag gac cta aac ttg cct gct gtt ctc agc gtg 912Ala His Tyr Cys Glu Lys Asp Leu Asn Leu Pro Ala Val Leu Ser Val 290 295 300gca aac cag atc aca cgc gct ctg ctc ggt gtg gaa gcc cac gag atc 960Ala Asn Gln Ile Thr Arg Ala Leu Leu Gly Val Glu Ala His Glu Ile305 310 315 320agc atg ctt ttt ctc acc gac tac atc aag agt gcc acc ggt ctc agt 1008Ser Met Leu Phe Leu Thr Asp Tyr Ile Lys Ser Ala Thr Gly Leu Ser 325 330 335aat att gtc tcg gac aag aaa gac ggc ggg cag tat atg cga tgc aaa 1056Asn Ile Val Ser Asp Lys Lys Asp Gly Gly Gln Tyr Met Arg Cys Lys 340 345 350aca ggt atg cag tcg att tgc cat gcc atg tca aag gaa ctt gtt cca 1104Thr Gly Met Gln Ser Ile Cys His Ala Met Ser Lys Glu Leu Val Pro 355 360 365ggc tca gtg cac ctc aac acc ccc gtc gct gga att gag cag tcg gcg 1152Gly Ser Val His Leu Asn Thr Pro Val Ala Gly Ile Glu Gln Ser Ala 370 375 380tcc ggc tgt ata gta cga tcg gcc tcg ggc gcc gtg ttc cga agc aaa 1200Ser Gly Cys Ile Val Arg Ser Ala Ser Gly Ala Val Phe Arg Ser Lys385 390 395 400aag gtg gtg gtt tcg tta ccg aca aca ttg tat ccc acc ttg aca ttt 1248Lys Val Val Val Ser Leu Pro Thr Thr Leu Tyr Pro Thr Leu Thr Phe 405 410 415tca cca cct ctt ccc gcc gag aag caa gca ttg gcg gaa aaa tct atc 1296Ser Pro Pro Leu Pro Ala Glu Lys Gln Ala Leu Ala Glu Lys Ser Ile 420 425 430ctc ggc tac tat agc aag ata gtc ttc gta tgg gac aac ccg tgg tgg 1344Leu Gly Tyr Tyr Ser Lys Ile Val Phe Val Trp Asp Asn Pro Trp Trp 435 440 445cgc gaa caa ggc ttc tcg ggc gtc ctc caa tcg agc tgt gac ccc atc 1392Arg Glu Gln Gly Phe Ser Gly Val Leu Gln Ser Ser Cys Asp Pro Ile 450 455 460tca ttt gcc aga gat acc agc atc gaa gtc gat cgg caa tgg tcc att 1440Ser Phe Ala Arg Asp Thr Ser Ile Glu Val Asp Arg Gln Trp Ser Ile465 470 475 480acc tgt ttc atg gtc gga gac ccg gga cgg aag tgg tcc caa cag tcc 1488Thr Cys Phe Met Val Gly Asp Pro Gly Arg Lys Trp Ser Gln Gln Ser 485 490 495aag cag gta cga caa aag tct gtc tgg gac caa ctc cgc gca gcc tac 1536Lys Gln Val Arg Gln Lys Ser Val Trp Asp Gln Leu Arg Ala Ala Tyr 500 505 510gag aac gcc ggg gcc caa gtc cca gag ccg gcc aac gtg ctc gaa atc 1584Glu Asn Ala Gly Ala Gln Val Pro Glu Pro Ala Asn Val Leu Glu Ile 515 520 525gag tgg tcg aag cag cag tat ttc caa gga gct ccg agc gcc gtc tat 1632Glu Trp Ser Lys Gln Gln Tyr Phe Gln Gly Ala Pro Ser Ala Val Tyr 530 535 540ggg ctg aac gat ctc atc aca ctg ggt tcg gcg ctc aga acg ccg ttc 1680Gly Leu Asn Asp Leu Ile Thr Leu Gly Ser Ala Leu Arg Thr Pro Phe545 550 555 560aag tgt gtt cat ttc gtt gga acg gag acg tct tta gtt tgg aaa ggg 1728Lys Cys Val His Phe Val Gly Thr Glu Thr Ser Leu Val Trp Lys Gly 565 570 575tat atg gaa ggg gcc ata cga tcg ggt caa cga ggt gct gca gaa gtt 1776Tyr Met Glu Gly Ala Ile Arg Ser Gly Gln Arg Gly Ala Ala Glu Val 580 585 590gtg gct agc ctg gtg cca gca gca tag 1803Val Ala Ser Leu Val Pro Ala Ala * 595 600 25 600 PRT Exophiala spinifera 25Met Ala Leu Ala Pro Ser Tyr Ile Asn Pro Pro Asn Val Ala Ser Pro 1 5 10 15Ala Gly Tyr Ser His Ile Gly Val Gly Pro Asn Glu Ala Arg Tyr Val 20 25 30Thr Ile Ala Gly Gln Ile Gly Gln Asp Ala Leu Gly Val Thr Asp Pro 35 40 45Ala Tyr Glu Lys Gln Val Ala Gln Ala Phe Ala Asn Leu Arg Ala Cys 50 55 60Leu Ala Ala Val Gly Ala Ser Ser Asn Asp Val Thr Lys Leu Asn Tyr65 70 75 80Tyr Ile Val Asp Tyr Ala Pro Ser Lys Leu Thr Ala Ile Gly Asp Gly 85 90 95Leu Lys Ser Thr Phe Ala Leu Asp Arg Leu Pro Pro Cys Thr Leu Val 100 105 110Pro Val Pro Ala Leu Ala Ser Pro Glu Tyr Leu Phe Glu Val Asp Ala 115 120 125Thr Ala Leu Val Pro Gly His Ser Thr Pro Asp Asn Val Ala Asp Val 130 135 140Val Val Val Gly Ala Gly Leu Ser Gly Leu Glu Thr Ala Arg Lys Val145 150 155 160Gln Ala Ala Gly Leu Ser Cys Leu Val Leu Glu Ala Met Asp Arg Val 165 170 175Gly Gly Lys Thr Leu Ser Val Gln Ser Gly Pro Gly Arg Thr Thr Ile 180 185 190Asn Asp Leu Gly Ala Ala Trp Ile Asn Asp Ser Asn Gln Ser Glu Val 195 200 205Ser Arg Leu Phe Glu Arg Phe His Leu Glu Gly Glu Leu Gln Arg Thr 210 215 220Thr Gly Asn Ser Ile His Gln Ala Gln Asp Gly Thr Thr Thr Thr Ala225 230 235 240Pro Tyr Gly Asp Ser Pro Leu Ser Glu Glu Val Ala Ser Ala Leu Ala 245 250 255Glu Leu Leu Pro Val Trp Ser Gln Leu Ile Glu Glu Tyr Ser Leu Glu 260 265 270Asp Pro Lys Ala Ser Pro Gln Ala Lys Arg Leu Asp Ser Val Ser Phe 275 280 285Ala His Tyr Cys Glu Lys Asp Leu Asn Leu Pro Ala Val Leu Ser Val 290 295 300Ala Asn Gln Ile Thr Arg Ala Leu Leu Gly Val Glu Ala His Glu Ile305 310 315 320Ser Met Leu Phe Leu Thr Asp Tyr Ile Lys Ser Ala Thr Gly Leu Ser 325 330 335Asn Ile Val Ser Asp Lys Lys Asp Gly Gly Gln Tyr Met Arg Cys Lys 340 345 350Thr Gly Met Gln Ser Ile Cys His Ala Met Ser Lys Glu Leu Val Pro 355 360 365Gly Ser Val His Leu Asn Thr Pro Val Ala Gly Ile Glu Gln Ser Ala 370 375 380Ser Gly Cys Ile Val Arg Ser Ala Ser Gly Ala Val Phe Arg Ser Lys385 390 395 400Lys Val Val Val Ser Leu Pro Thr Thr Leu Tyr Pro Thr Leu Thr Phe 405 410 415Ser Pro Pro Leu Pro Ala Glu Lys Gln Ala Leu Ala Glu Lys Ser Ile 420 425 430Leu Gly Tyr Tyr Ser Lys Ile Val Phe Val Trp Asp Asn Pro Trp Trp 435 440 445Arg Glu Gln Gly Phe Ser Gly Val Leu Gln Ser Ser Cys Asp Pro Ile 450 455 460Ser Phe Ala Arg Asp Thr Ser Ile Glu Val Asp Arg Gln Trp Ser Ile465 470 475 480Thr Cys Phe Met Val Gly Asp Pro Gly Arg Lys Trp Ser Gln Gln Ser 485 490 495Lys Gln Val Arg Gln Lys Ser Val Trp Asp Gln Leu Arg Ala Ala Tyr 500 505 510Glu Asn Ala Gly Ala Gln Val Pro Glu Pro Ala Asn Val Leu Glu Ile 515 520 525Glu Trp Ser Lys Gln Gln Tyr Phe Gln Gly Ala Pro Ser Ala Val Tyr 530 535 540Gly Leu Asn Asp Leu Ile Thr Leu Gly Ser Ala Leu Arg Thr Pro Phe545 550 555 560Lys Cys Val His Phe Val Gly Thr Glu Thr Ser Leu Val Trp Lys Gly 565 570 575Tyr Met Glu Gly Ala Ile Arg Ser Gly Gln Arg Gly Ala Ala Glu Val 580 585 590Val Ala Ser Leu Val Pro Ala Ala 595 600 26 1803 DNA Exophiala spinifera CDS (1)...(1803) misc_feature (0)...(0) isolate ESP002_C12 26atg gca ctt gca ccg agc tac atc aat ccc cca aac gtc gcc tcc cca 48Met Ala Leu Ala Pro Ser Tyr Ile Asn Pro Pro Asn Val Ala Ser Pro 1 5 10 15gca ggg tat tct cac gtc ggc gta ggc cca gac gga ggg agg tat gtg 96Ala Gly Tyr Ser His Val Gly Val Gly Pro Asp Gly Gly Arg Tyr Val 20 25 30aca ata gct gga cag att gga caa gac gct tcg ggc gtg aca gac cct 144Thr Ile Ala Gly Gln Ile Gly Gln Asp Ala Ser Gly Val Thr Asp Pro 35 40 45gcc tac gag aaa cag gtt gcc caa gca ttc gcc aat ctg cga gct tgc 192Ala Tyr Glu Lys Gln Val Ala Gln Ala Phe Ala Asn Leu Arg Ala Cys 50 55 60ctt gct gca gtt gga gcc act tca aac gac gtc acc aag ctc aat tac 240Leu Ala Ala Val Gly Ala Thr Ser Asn Asp Val Thr Lys Leu Asn Tyr 65 70 75 80tac atc gtc gac tac gcc ccg agc aaa ctc acc gca att gga gat ggg 288Tyr Ile Val Asp Tyr Ala Pro Ser Lys Leu Thr Ala Ile Gly Asp Gly 85 90 95ctg aag gct acc ttt gcc ctt gac agg ctc cct cct tgc acg ctg gtg 336Leu Lys Ala Thr Phe Ala Leu Asp Arg Leu Pro Pro Cys Thr Leu Val 100 105 110cca gtg tcg gcc ttg tct tca cct gaa tac ctc ttt gag gtt gat gcc 384Pro Val Ser Ala Leu Ser Ser Pro Glu Tyr Leu Phe Glu Val Asp Ala 115 120 125acg gcg ctg gtg ccg gga cac acg acc cca gac aac gtt gcg gac gtg 432Thr Ala Leu Val Pro Gly His Thr Thr Pro Asp Asn Val Ala Asp Val 130 135 140gta gtg gtg ggc gct ggc ttg agc ggt ttg gag acg gca cgc aaa gtc 480Val Val Val Gly Ala Gly Leu Ser Gly Leu Glu Thr Ala Arg Lys Val145 150 155 160cag gcc gcc ggt ctg tcc tgc ctc gtt ctt gag gcg atg gat cgt gta 528Gln Ala Ala Gly Leu Ser Cys Leu Val Leu Glu Ala Met Asp Arg Val 165 170 175ggg gga aag act ctg agc gta caa tcg ggt ccc ggc agg acg act atc 576Gly Gly Lys Thr Leu Ser Val Gln Ser Gly Pro Gly Arg Thr Thr Ile 180 185 190aac gac ctc ggc gct gcg tgg atc aat gac agc aac caa agc gaa gta 624Asn Asp Leu Gly Ala Ala Trp Ile Asn Asp Ser Asn Gln Ser Glu Val 195 200 205tcc aga ttg ttt gaa aga ttt cat ttg gag ggc gag ctc cag agg acg 672Ser Arg Leu Phe Glu Arg Phe His Leu Glu Gly Glu Leu Gln Arg Thr 210 215 220act gga aat tca atc cat caa gca caa gac ggt aca acc act aca gct 720Thr Gly Asn Ser Ile His Gln Ala Gln Asp Gly Thr Thr Thr Thr Ala225 230 235 240cct tat ggt gac tcc ttg ctg agc gag gag gtt gca agt gca ctt gcg 768Pro Tyr Gly Asp Ser Leu Leu Ser Glu Glu Val Ala Ser Ala Leu Ala 245 250 255gaa ctc ctc ccc gta tgg tct cag ctg atc gaa gag cat agc ctt caa 816Glu Leu Leu Pro Val Trp Ser Gln Leu Ile Glu Glu His Ser Leu Gln 260 265 270gac ctc aag gcg agc cct cag gcg aag cgg ctc gac agt gtg agc ttc 864Asp Leu Lys Ala Ser Pro Gln Ala Lys Arg Leu Asp Ser Val Ser Phe 275 280 285gcg cac tac tgt gag aag gaa cta aac ttg cct gct gtt ctc ggc gta 912Ala His Tyr Cys Glu Lys Glu Leu Asn Leu Pro Ala Val Leu Gly Val 290 295 300gca aac cag atc aca cgc gct ctg ctc ggt gtg gaa gcc cac gag atc 960Ala Asn Gln Ile Thr Arg Ala Leu Leu Gly Val Glu Ala His Glu Ile305 310 315 320agc atg ctt ttt ctc acc gac tac atc aag agt gcc acc ggt ctc agt 1008Ser Met Leu Phe Leu Thr Asp Tyr Ile Lys Ser Ala Thr Gly Leu Ser 325 330 335aat att ttc tcg gac aag aaa gac ggc ggg cag tat atg cga tgc aaa 1056Asn Ile Phe Ser Asp Lys Lys Asp Gly Gly Gln Tyr Met Arg Cys Lys 340 345 350aca ggt atg cag tcg att tgc cat gcc atg tca aag gaa ctt gtt cca 1104Thr Gly Met Gln Ser Ile Cys His Ala Met Ser Lys Glu Leu Val Pro 355 360 365ggc tca gtg cac ctc aac acc ccc gtc gct gaa att gag cag tcg gca 1152Gly Ser Val His Leu Asn Thr Pro Val Ala Glu Ile Glu Gln Ser Ala 370 375 380tcc ggc tgt aca gta cga tcg gcc tcg ggc gcc gtg ttc cga agc aaa 1200Ser Gly Cys Thr Val Arg Ser Ala Ser Gly Ala Val Phe Arg Ser Lys385 390 395 400aag gtg gtg gtt tcg tta ccg aca acc ttg tat ccc acc ttg aca ttt 1248Lys Val Val Val Ser Leu Pro Thr Thr Leu Tyr Pro Thr Leu Thr Phe 405 410 415tca cca cct ctc ccc gcc gag aag caa gca ttg gcg gaa aat tct atc 1296Ser Pro Pro Leu Pro Ala Glu Lys Gln Ala Leu Ala Glu Asn Ser Ile 420 425 430ctg ggc tac tat agc aag ata gtc ttc gta tgg gac aag ccg tgg tgg 1344Leu Gly Tyr Tyr Ser Lys Ile Val Phe Val Trp Asp Lys Pro Trp Trp 435 440 445cgc gaa caa ggc ttc tcg ggc gtc ctc caa tcg agc tgt gac ccc atc 1392Arg Glu Gln Gly Phe Ser Gly Val Leu Gln Ser Ser Cys Asp Pro Ile 450 455 460tca ttt gcc aga gat acc agc atc gac gtc gat cga caa tgg tcc att 1440Ser Phe Ala Arg Asp Thr Ser Ile Asp Val Asp Arg Gln Trp Ser Ile465 470 475 480acc tgt ttc atg gtc gga gac ccg gga cgg aag tgg tcc caa cag tcc 1488Thr Cys Phe Met Val Gly Asp Pro Gly Arg Lys Trp Ser Gln Gln Ser 485 490 495aag cag gta cga caa aag tct gtc tgg gac caa ctc cgc gca gcc tac 1536Lys Gln Val Arg Gln Lys Ser Val Trp Asp Gln Leu Arg Ala Ala Tyr 500 505 510gag aac gcc ggg gcc caa gtc cca gag ccg gcc aac gtg ctc gaa atc 1584Glu Asn Ala Gly Ala Gln Val Pro Glu Pro Ala Asn Val Leu Glu Ile 515 520 525gag tgg tcg aag cag cag tat ttc caa gga gct ccg agc gcc gtc tat 1632Glu Trp Ser Lys Gln Gln Tyr Phe Gln Gly Ala Pro Ser Ala Val Tyr 530 535 540ggg ctg aac gat ctc atc aca ctg ggt tcg gcg ctc aga acg ccg ttc 1680Gly Leu Asn Asp Leu Ile Thr Leu Gly Ser Ala Leu Arg Thr Pro Phe545 550 555 560aag agt gtt cat ttc gtt gga acg gag acg tct tta gtt tgg aaa ggg 1728Lys Ser Val His Phe Val Gly Thr Glu Thr Ser Leu Val Trp Lys Gly 565 570 575tat atg gaa ggg gcc ata cga tcg ggt caa cga ggt gct gca gaa gtt 1776Tyr Met Glu Gly Ala Ile Arg Ser Gly Gln Arg Gly Ala Ala Glu Val 580 585 590gtg gct agc ctg gtg cca gca gca tag 1803Val Ala Ser Leu Val Pro Ala Ala * 595 600 27 600 PRT Exophiala spinifera 27Met Ala Leu Ala Pro Ser Tyr Ile Asn Pro Pro Asn Val Ala Ser Pro 1 5 10 15Ala Gly Tyr Ser His Val Gly Val Gly Pro Asp Gly Gly Arg Tyr Val 20 25 30Thr Ile Ala Gly Gln Ile Gly Gln Asp Ala Ser Gly Val Thr Asp Pro 35 40 45Ala Tyr Glu Lys Gln Val Ala Gln Ala Phe Ala Asn Leu Arg Ala Cys 50 55 60Leu Ala Ala Val Gly Ala Thr Ser Asn Asp Val Thr Lys Leu Asn Tyr65 70 75 80Tyr Ile Val Asp Tyr Ala Pro Ser Lys Leu Thr Ala Ile Gly Asp Gly 85 90 95Leu Lys Ala Thr Phe Ala Leu Asp Arg Leu Pro Pro Cys Thr Leu Val 100 105 110Pro Val Ser Ala Leu Ser Ser Pro Glu Tyr Leu Phe Glu Val Asp Ala 115 120 125Thr Ala Leu Val Pro Gly His Thr Thr Pro Asp Asn Val Ala Asp Val 130 135 140Val Val Val Gly Ala Gly Leu Ser Gly Leu Glu Thr Ala Arg Lys Val145 150 155 160Gln Ala Ala Gly Leu Ser Cys Leu Val Leu Glu Ala Met Asp Arg Val 165 170 175Gly Gly Lys Thr Leu Ser Val Gln Ser Gly Pro Gly Arg Thr Thr Ile 180 185 190Asn Asp Leu Gly Ala Ala Trp Ile Asn Asp Ser Asn Gln Ser Glu Val 195 200 205Ser Arg Leu Phe Glu Arg Phe His Leu Glu Gly Glu Leu Gln Arg Thr 210 215 220Thr Gly Asn Ser Ile His Gln Ala Gln Asp Gly Thr Thr Thr Thr Ala225 230 235 240Pro Tyr Gly Asp Ser Leu Leu Ser Glu Glu Val Ala Ser Ala Leu Ala 245 250 255Glu Leu Leu Pro Val Trp Ser Gln Leu Ile Glu Glu His Ser Leu Gln 260 265 270Asp Leu Lys Ala Ser Pro Gln Ala Lys Arg Leu Asp Ser Val Ser Phe 275 280 285Ala His Tyr Cys Glu Lys Glu Leu Asn Leu Pro Ala Val Leu Gly Val 290 295 300Ala Asn Gln Ile Thr Arg Ala Leu Leu Gly Val Glu Ala His Glu Ile305 310 315 320Ser Met Leu Phe Leu Thr Asp Tyr Ile Lys Ser Ala Thr Gly Leu Ser 325 330 335Asn Ile Phe Ser Asp Lys Lys Asp Gly Gly Gln Tyr Met Arg Cys Lys 340 345 350Thr Gly Met Gln Ser Ile Cys His Ala Met Ser Lys Glu Leu Val Pro 355 360 365Gly Ser Val His Leu Asn Thr Pro Val Ala Glu Ile Glu Gln Ser Ala 370 375 380Ser Gly Cys Thr Val Arg Ser Ala Ser Gly Ala Val Phe Arg Ser Lys385 390 395 400Lys Val Val Val Ser Leu Pro Thr Thr Leu Tyr Pro Thr Leu Thr Phe 405 410 415Ser Pro Pro Leu Pro Ala Glu Lys Gln Ala Leu Ala Glu Asn Ser Ile 420 425 430Leu Gly Tyr Tyr Ser Lys Ile Val Phe Val Trp Asp Lys Pro Trp Trp 435 440 445Arg Glu Gln Gly Phe Ser Gly Val Leu Gln Ser Ser Cys Asp Pro Ile 450 455 460Ser Phe Ala Arg Asp Thr Ser Ile Asp Val Asp Arg Gln Trp Ser Ile465 470 475 480Thr Cys Phe Met Val Gly Asp Pro Gly Arg Lys Trp Ser Gln Gln Ser 485 490 495Lys Gln Val Arg Gln Lys Ser Val Trp Asp Gln Leu Arg Ala Ala Tyr 500 505 510Glu Asn Ala Gly Ala Gln Val Pro Glu Pro Ala Asn Val Leu Glu Ile 515 520 525Glu Trp Ser Lys Gln Gln Tyr Phe Gln Gly Ala Pro Ser Ala Val Tyr 530 535 540Gly Leu Asn Asp Leu Ile Thr Leu Gly Ser Ala Leu Arg Thr Pro Phe545 550 555 560Lys Ser Val His Phe Val Gly Thr Glu Thr Ser Leu Val Trp Lys Gly 565 570 575Tyr Met Glu Gly Ala Ile Arg Ser Gly Gln Arg Gly Ala Ala Glu Val 580 585 590Val Ala Ser Leu Val Pro Ala Ala 595 600 28 1803 DNA Rhinocladiella atrovirens CDS (1)...(1803) misc_feature (0)...(0) isolate C1 28atg gca ctt gca ccg agc tac atc aat ccc cca aac ctc gcc tcc cca 48Met Ala Leu Ala Pro Ser Tyr Ile Asn Pro Pro Asn Leu Ala Ser Pro 1 5 10 15gca ggg tat tcc cac gtc ggc gta ggc cca aac gga ggg agg tat gcg 96Ala Gly Tyr Ser His Val Gly Val Gly Pro Asn Gly Gly Arg Tyr Ala 20 25 30aca ata gct gga cag att gga caa gac gct tcg gcc gtg aca gac cct 144Thr Ile Ala Gly Gln Ile Gly Gln Asp Ala Ser Ala Val Thr Asp Pro 35 40 45gcc tac gag aaa cag gtt gcc caa gca ttc gcc aac ctg cga gct tgt 192Ala Tyr Glu Lys Gln Val Ala Gln Ala Phe Ala Asn Leu Arg Ala Cys 50 55 60ctt gct gca gtt gga gcc act tca aac gac att acc aag ctc aat tac 240Leu Ala Ala Val Gly Ala Thr Ser Asn Asp Ile Thr Lys Leu Asn Tyr 65 70 75 80tac atc gtc gac tac aac ccg agc aaa ctc acc gca att gga gat ggg 288Tyr Ile Val Asp Tyr Asn Pro Ser Lys Leu Thr Ala Ile Gly Asp Gly 85 90 95ctg aag gct acc ttt gcc ctt gac agg ctc cct cct tgc acg ctg gtg 336Leu Lys Ala Thr Phe Ala Leu Asp Arg Leu Pro Pro Cys Thr Leu Val 100 105 110cca gtg ccg gcc ctg gct tca cct gaa tac ccc ttt gag gtt gat gcc 384Pro Val Pro Ala Leu Ala Ser Pro Glu Tyr Pro Phe Glu Val Asp Ala 115 120 125acg gcg ctg gtt cca gga cac tca acc cca gac aat gtt gcg gac gtg 432Thr Ala Leu Val Pro Gly His Ser Thr Pro Asp Asn Val Ala Asp Val 130 135 140gtc gtg gtg ggc gct ggc ttg agc ggt ttg gag acg gca cgc aaa gtc 480Val Val Val Gly Ala Gly Leu Ser Gly Leu Glu Thr Ala Arg Lys Val145 150 155 160cag gct gcc ggg ctg tcc tgc ctc gtt ctt gag gcg atg gat cgt gtg 528Gln Ala Ala Gly Leu Ser Cys Leu Val Leu Glu Ala Met Asp Arg Val 165 170 175ggg gga aag act ctg agc gta caa tcg ggt ccc ggc agg acg gct atc 576Gly Gly Lys Thr Leu Ser Val Gln Ser Gly Pro Gly Arg Thr Ala Ile 180 185 190aat gac ctc ggc gct gcg tgg atc aat gac agc aac caa agc gaa gta 624Asn Asp Leu Gly Ala Ala Trp Ile Asn Asp Ser Asn Gln Ser Glu Val 195 200 205ttc aaa tta ttt gaa aga ttt cat ttg gag ggc gag ctc cag agg acg 672Phe Lys Leu Phe Glu Arg Phe His Leu Glu Gly Glu Leu Gln Arg Thr 210 215 220acc gga aat tca atc cat caa gca caa gac ggt aca acc act aca gct 720Thr Gly Asn Ser Ile His Gln Ala Gln Asp Gly Thr Thr Thr Thr Ala225 230 235 240cct tat ggt gat tcc ctg ctg agc gag gag gtt gca agt gca ctc gcg 768Pro Tyr Gly Asp Ser Leu Leu Ser Glu Glu Val Ala Ser Ala Leu Ala 245 250 255gaa ctc ctt ccc gca tgg tct cag ctg atc gaa gag cat agt ctt gaa 816Glu Leu Leu Pro Ala Trp Ser Gln Leu Ile Glu Glu His Ser Leu Glu 260 265 270gac ccc aag gcg agc cct caa gcg aag cag ctc gac agt gtg agc ttc 864Asp Pro Lys Ala Ser Pro Gln Ala Lys Gln Leu Asp Ser Val Ser Phe 275 280 285gca cac tac tgt gag aag gat cta agc ttg cct gct gtt ctc ggc gtg 912Ala His Tyr Cys Glu Lys Asp Leu Ser Leu Pro Ala Val Leu Gly Val 290 295 300gca aac cag atc aca cgc gct ctg ctc ggt gtg gaa gcc cac gag atc 960Ala Asn Gln Ile Thr Arg Ala Leu Leu Gly Val Glu Ala His Glu Ile305 310 315 320agc atg ctt ttt ctc acc gac tac atc aag agt gcc acc ggt ctc agt 1008Ser Met Leu Phe Leu Thr Asp Tyr Ile Lys Ser Ala Thr Gly Leu Ser 325 330 335aat att gtc tcg gat aag aaa gac ggt ggg cag tat atg cga tgc aaa 1056Asn Ile Val Ser Asp Lys Lys Asp Gly Gly Gln Tyr Met Arg Cys Lys 340 345 350aca ggt atg cag tcg ctt tgc cat gcc atg tca aag gaa ctt gtt cca 1104Thr Gly Met Gln Ser Leu Cys His Ala Met Ser Lys Glu Leu Val Pro 355 360 365ggc tca gtg cac ctc aac acc ccc gtc gcc gaa att gag cag tcg gca 1152Gly Ser Val His Leu Asn Thr Pro Val Ala Glu Ile Glu Gln Ser Ala 370 375 380tcc ggc tgt aca gta cga tcg gcc tcg ggc ggc gtg ttc cga agt aaa 1200Ser Gly Cys Thr Val Arg Ser Ala Ser Gly Gly Val Phe Arg Ser Lys385 390 395 400aag gtg gtg gtt tcg tta ccg aca acc ttg tat ccc acc ttg ata ttt 1248Lys Val Val Val Ser Leu Pro Thr Thr Leu Tyr Pro Thr Leu Ile Phe 405 410 415tca cca cct ctt ccc gcc gag aag caa gca ttg gct gaa aaa tcc atc 1296Ser Pro Pro Leu Pro Ala Glu Lys Gln Ala Leu Ala Glu Lys Ser Ile 420 425 430ctg ggc tac tat agc aag ata gtc ttc gta tgg gac aag ccg tgg tgg 1344Leu Gly Tyr Tyr Ser Lys Ile Val Phe Val Trp Asp Lys Pro Trp Trp 435 440 445cgc gaa caa ggc ttc tcg ggc gtc ctc caa tcg agc tgt gac ccc atc 1392Arg Glu Gln Gly Phe Ser Gly Val Leu Gln Ser Ser Cys Asp Pro Ile 450 455 460tca ttt gcc aga gat acc agc atc gaa gtc gat cgg caa tgg tcc att 1440Ser Phe Ala Arg Asp Thr Ser Ile Glu Val Asp Arg Gln Trp Ser Ile465 470 475 480acc tgt ttc atg gtc gga gac ccg gga cgg aag tgg tcc caa cag tcc 1488Thr Cys Phe Met Val Gly Asp Pro Gly Arg Lys Trp Ser Gln Gln Ser 485 490 495aag cag gta cga cag aag tct gtc tgg aac caa ctc cgc gca gcc tac 1536Lys Gln Val Arg Gln Lys Ser Val Trp Asn Gln Leu Arg Ala Ala Tyr 500 505 510gag aac gcc ggg gcc caa gtc cca gag ccg gcc aac gtg ctc gag atc 1584Glu Asn Ala Gly Ala Gln Val Pro Glu Pro Ala Asn Val Leu Glu Ile 515 520 525gag tgg tcg aag cag cag tat ttc caa gga gcg ccg agc gtc gtc tat 1632Glu Trp Ser Lys Gln Gln Tyr Phe Gln Gly Ala Pro Ser Val Val Tyr 530 535 540ggg ctg aac tgt ctc aac aca ctg ggt tcg gcg ctc aga acg ccg ttc 1680Gly Leu Asn Cys Leu Asn Thr Leu Gly Ser Ala Leu Arg Thr Pro Phe545 550 555 560aag ggt gtt cat ttc gtt gga acg gag acg tct ttg gtt tgg aaa ggg 1728Lys Gly Val His Phe Val Gly Thr Glu Thr Ser Leu Val Trp Lys Gly 565 570 575tat atg gaa ggg gcc ata cga tcg ggt cag cga ggc gct gca gaa gtt 1776Tyr Met Glu Gly Ala Ile Arg Ser Gly Gln Arg Gly Ala Ala Glu Val 580 585 590gtg gct agc ctg gtg cca gca gca tag 1803Val Ala Ser Leu Val Pro Ala Ala * 595 600 29 600 PRT Rhinocladiella atrovirens 29Met Ala Leu Ala Pro Ser Tyr Ile Asn Pro Pro Asn Leu Ala Ser Pro 1 5 10 15Ala Gly Tyr Ser His Val Gly Val Gly Pro Asn Gly Gly Arg Tyr Ala 20 25 30Thr Ile Ala Gly Gln Ile Gly Gln Asp Ala Ser Ala Val Thr Asp Pro 35 40 45Ala Tyr Glu Lys Gln Val Ala Gln Ala Phe Ala Asn Leu Arg Ala Cys 50 55 60Leu Ala Ala Val Gly Ala Thr Ser Asn Asp Ile Thr Lys Leu Asn Tyr65 70 75 80Tyr Ile Val Asp Tyr Asn Pro Ser Lys Leu Thr Ala Ile Gly Asp Gly 85 90 95Leu Lys Ala Thr Phe Ala Leu Asp Arg Leu Pro Pro Cys Thr Leu Val 100 105 110Pro Val Pro Ala Leu Ala Ser Pro Glu Tyr Pro Phe Glu Val Asp Ala 115 120 125Thr Ala Leu Val Pro Gly His Ser Thr Pro Asp Asn Val Ala Asp Val 130 135 140Val Val Val Gly Ala Gly Leu Ser Gly Leu Glu Thr Ala Arg Lys Val145 150 155 160Gln Ala Ala Gly Leu Ser Cys Leu Val Leu Glu Ala Met Asp Arg Val 165 170 175Gly Gly Lys Thr Leu Ser Val Gln Ser Gly Pro Gly Arg Thr Ala Ile 180 185 190Asn Asp Leu Gly Ala Ala Trp Ile Asn Asp Ser Asn Gln Ser Glu Val 195 200 205Phe Lys Leu Phe Glu Arg Phe His Leu Glu Gly Glu Leu Gln Arg Thr 210 215 220Thr Gly Asn Ser Ile His Gln Ala Gln Asp Gly Thr Thr Thr Thr Ala225 230 235 240Pro Tyr Gly Asp Ser Leu Leu Ser Glu Glu Val Ala Ser Ala Leu Ala 245 250 255Glu Leu Leu Pro Ala Trp Ser Gln Leu Ile Glu Glu His Ser Leu Glu 260 265 270Asp Pro Lys Ala Ser Pro Gln Ala Lys Gln Leu Asp Ser Val Ser Phe 275 280 285Ala His Tyr Cys Glu Lys Asp Leu Ser Leu Pro Ala Val Leu Gly Val 290 295 300Ala Asn Gln Ile Thr Arg Ala Leu Leu Gly Val Glu Ala His Glu Ile305 310 315 320Ser Met Leu Phe Leu Thr Asp Tyr Ile Lys Ser Ala Thr Gly Leu Ser 325 330 335Asn Ile Val Ser Asp Lys Lys Asp Gly Gly Gln Tyr Met Arg Cys Lys 340 345 350Thr Gly Met Gln Ser Leu Cys His Ala Met Ser Lys Glu Leu Val Pro 355 360 365Gly Ser Val His Leu Asn Thr Pro Val Ala Glu Ile Glu Gln Ser Ala 370 375 380Ser Gly Cys Thr Val Arg Ser Ala Ser Gly Gly Val Phe Arg Ser Lys385 390 395 400Lys Val Val Val Ser Leu Pro Thr Thr Leu Tyr Pro Thr Leu Ile Phe 405 410 415Ser Pro Pro Leu Pro Ala Glu Lys Gln Ala Leu Ala Glu Lys Ser Ile 420 425 430Leu Gly Tyr Tyr Ser Lys Ile Val Phe Val Trp Asp Lys Pro Trp Trp 435 440 445Arg Glu Gln Gly Phe Ser Gly Val Leu Gln Ser Ser Cys Asp Pro Ile 450 455 460Ser Phe Ala Arg Asp Thr Ser Ile Glu Val Asp Arg Gln Trp Ser Ile465 470 475 480Thr Cys Phe Met Val Gly Asp Pro Gly Arg Lys Trp Ser Gln Gln Ser 485 490 495Lys Gln Val Arg Gln Lys Ser Val Trp Asn Gln Leu Arg Ala Ala Tyr 500 505 510Glu Asn Ala Gly Ala Gln Val Pro Glu Pro Ala Asn Val Leu Glu Ile 515 520 525Glu Trp Ser Lys Gln Gln Tyr Phe Gln Gly Ala Pro Ser Val Val Tyr 530 535 540Gly Leu Asn Cys Leu Asn Thr Leu Gly Ser Ala Leu Arg Thr Pro Phe545 550 555 560Lys Gly Val His Phe Val Gly Thr Glu Thr Ser Leu Val Trp Lys Gly 565 570 575Tyr Met Glu Gly Ala Ile Arg Ser Gly Gln Arg Gly Ala Ala Glu Val 580 585 590Val Ala Ser Leu Val Pro Ala Ala 595 600 30 1803 DNA Rhinocladiella atrovirens CDS (1)...(1803) misc_feature (0)...(0) isolate C2 30atg gca ctt gca ccg agc tac atc aat ccc cca aac ctc gcc tcc cca 48Met Ala Leu Ala Pro Ser Tyr Ile Asn Pro Pro Asn Leu Ala Ser Pro 1 5 10 15gca ggg tat tcc tac gtc ggc gta ggc cca aac gga ggg agg tat gtg 96Ala Gly Tyr Ser Tyr Val Gly Val Gly Pro Asn Gly Gly Arg Tyr Val 20 25 30aca ata gct gga cag att gga caa gac gct tcg gcc gtg aca gac cct 144Thr Ile Ala Gly Gln Ile Gly Gln Asp Ala Ser Ala Val Thr Asp Pro 35 40 45gcc tac gag aaa cag gtt gcc caa gca ttc gcc aac ctg cga gct tgt 192Ala Tyr Glu Lys Gln Val Ala Gln Ala Phe Ala Asn Leu Arg Ala Cys 50 55 60ctt gct gca gtt gga gcc act tca aac gac att acc aag ctc aat tac 240Leu Ala Ala Val Gly Ala Thr Ser Asn Asp Ile Thr Lys Leu Asn Tyr 65 70 75 80tac atc gtc gac tac aac ccg agc aaa ctc acc gca att gga gat ggg 288Tyr Ile Val Asp Tyr Asn Pro Ser Lys Leu Thr Ala Ile Gly Asp Gly 85 90 95ctg aag gct acc ttt gcc ctt gac agg ctc cct cct tgc acg ctg gtg 336Leu Lys Ala Thr Phe Ala Leu Asp Arg Leu Pro Pro Cys Thr Leu Val 100 105 110cca gtg ccg gcc ctg gct tca cct gaa tac ctc ttt gag gtt gat gcc 384Pro Val Pro Ala Leu Ala Ser Pro Glu Tyr Leu Phe Glu Val Asp Ala 115 120 125acg gcg ctg gtt cca gga cac tca acc cca gac aat gtt gcg gac gtg 432Thr Ala Leu Val Pro Gly His Ser Thr Pro Asp Asn Val Ala Asp Val 130 135 140gtc gtg gtg ggc gct ggc ttg agc ggt ttg gag acg gca cgc aaa gtc 480Val Val Val Gly Ala Gly Leu Ser Gly Leu Glu Thr Ala Arg Lys Val145 150 155 160cag gct gcc ggg ctg tcc tgc ctc gtt ctt gag gcg atg gat cgt gtg 528Gln Ala Ala Gly Leu Ser Cys Leu Val Leu Glu Ala Met Asp Arg Val 165 170 175ggg gga aag act ctg agc gta caa tcg ggt ccc ggc agg acg act atc 576Gly Gly Lys Thr Leu Ser Val Gln Ser Gly Pro Gly Arg Thr Thr Ile 180 185 190aat gac ctc ggc gct gcg tgg atc aat gac agc aac caa agc gaa gta 624Asn Asp Leu Gly Ala Ala Trp Ile Asn Asp Ser Asn Gln Ser Glu Val 195 200 205ttc aaa tta ttt gaa aga ttt cat ttg gag ggc gag ctc cag agg acg 672Phe Lys Leu Phe Glu Arg Phe His Leu Glu Gly Glu Leu Gln Arg Thr 210 215 220acc gga aat tca atc cat caa gca caa gac ggt aca acc act aca gct 720Thr Gly Asn Ser Ile His Gln Ala Gln Asp Gly Thr Thr Thr Thr Ala225 230 235 240cct tat ggt gat tcc ctg ctg agc gag gag gtt gca agt gca ctc gcg 768Pro Tyr Gly Asp Ser Leu Leu Ser Glu Glu Val Ala Ser Ala Leu Ala 245 250 255gaa ctc ctt ccc gca tgg tct cag ctg atc gaa gag cat agt ctt gaa 816Glu Leu Leu Pro Ala Trp Ser Gln Leu Ile Glu Glu His Ser Leu Glu 260 265 270gac ccc aag gcg agc cct caa gcg aag cag ctc gac agt gtg agc ttc 864Asp Pro Lys Ala Ser Pro Gln Ala Lys Gln Leu Asp Ser Val Ser Phe 275 280 285gca cac tac tgt gag aag gat cta aac ttg cct gct gtt ctc ggc gtg 912Ala His Tyr Cys Glu Lys Asp Leu Asn Leu Pro Ala Val Leu Gly Val 290 295 300gca aac cag atc aca cgc gct ctg ctc ggt gtg gaa gcc cac gag atc 960Ala Asn Gln Ile Thr Arg Ala Leu Leu Gly Val Glu Ala His Glu Ile305 310 315 320agc atg ttt ttt ctc acc gac tac atc aag agt gcc acc ggt ctc agt 1008Ser Met Phe Phe Leu Thr Asp Tyr Ile Lys Ser Ala Thr Gly Leu Ser 325 330 335aat att gtc tcg gat aag aaa gac ggt ggg cag tat atg cga tgc aaa 1056Asn Ile Val Ser Asp Lys Lys Asp Gly Gly Gln Tyr Met Arg Cys Lys 340 345 350aca ggt atg cag tcg ctt tgc cat gcc atg tca aag gaa ctt gtt cca 1104Thr Gly Met Gln Ser Leu Cys His Ala Met Ser Lys Glu Leu Val Pro 355 360 365ggc tca gtg cac ctc aac acc ccc gtc gcc gaa att gag cag tcg gca 1152Gly Ser Val His Leu Asn Thr Pro Val Ala Glu Ile Glu Gln Ser Ala 370 375 380tcc ggc tgt aca gta cga tcg gcc tcg ggc ggc gtg ttc cga agt aaa 1200Ser Gly Cys Thr Val Arg Ser Ala Ser Gly Gly Val Phe Arg Ser Lys385 390 395 400aag gtg gtg gtt tcg tta ccg aca acc ttg tat ccc acc ttg ata ttt 1248Lys Val Val Val Ser Leu Pro Thr Thr Leu Tyr Pro Thr Leu Ile Phe 405 410 415tca cca cct ctt ccc gcc gag aag caa gca ttg gct gaa aaa tcc atc 1296Ser Pro Pro Leu Pro Ala Glu Lys Gln Ala Leu Ala Glu Lys Ser Ile 420 425 430ctg ggc tac tat agc aag ata gtc ttc gta tgg gac aag ccg tgg tgg 1344Leu Gly Tyr Tyr Ser Lys Ile Val Phe Val Trp Asp Lys Pro Trp Trp 435 440 445cgc gaa caa ggc ttc tcg ggc gtc ctc caa tcg agc tgt gac ccc atc 1392Arg Glu Gln Gly Phe Ser Gly Val Leu Gln Ser Ser Cys Asp Pro Ile 450 455 460tca ttt gcc aga gat acc agc atc gaa gtc gat cgg caa tgg tcc att 1440Ser Phe Ala Arg Asp Thr Ser Ile Glu Val Asp Arg Gln Trp Ser Ile465 470 475 480acc tgt ttc atg gtc gga gac ccg gga cgg aag tgg tcc caa cag tcc 1488Thr Cys Phe Met Val Gly Asp Pro Gly Arg Lys Trp Ser Gln Gln Ser 485 490 495aag cag gta cga cag aag tct gtc tgg aac caa ctc cgc gca gcc tac 1536Lys Gln Val Arg Gln Lys Ser Val Trp Asn Gln Leu Arg Ala Ala Tyr 500 505 510gag aac gcc ggg gcc caa gtc cca gag ccg gcc aac gtg ctc gag atc 1584Glu Asn Ala Gly Ala Gln Val Pro Glu Pro Ala Asn Val Leu Glu Ile 515 520 525gag tgg tcg aag cag cag tat ttc caa gga gcg ccg agc gcc gtc tat 1632Glu Trp Ser Lys Gln Gln Tyr Phe Gln Gly Ala Pro Ser Ala Val Tyr 530 535 540ggg ctg aac tgt ctc aac aca ctg ggt tcg gcg ctc aga acg ccg ttc 1680Gly Leu Asn Cys Leu Asn Thr Leu Gly Ser Ala Leu Arg Thr Pro Phe545 550 555 560aag ggt gtt cat ttc gtt gga acg gag acg tct ttg gtt tgg aaa ggg 1728Lys Gly Val His Phe Val Gly Thr Glu Thr Ser Leu Val Trp Lys Gly 565 570 575tat atg gaa ggg gcc ata cga tcg ggt cag cga ggc gct gca gaa gtt 1776Tyr Met Glu Gly Ala Ile Arg Ser Gly Gln Arg Gly Ala Ala Glu Val 580 585 590gtg gct agc ctg gtg cca gca gca tag 1803Val Ala Ser Leu Val Pro Ala Ala * 595 600 31 600 PRT Rhinocladiella atrovirens 31Met Ala Leu Ala Pro Ser Tyr Ile Asn Pro Pro Asn Leu Ala Ser Pro 1 5 10 15Ala Gly Tyr Ser Tyr Val Gly Val Gly Pro Asn Gly Gly Arg Tyr Val 20 25 30Thr Ile Ala Gly Gln Ile Gly Gln Asp Ala Ser Ala Val Thr Asp Pro 35 40 45Ala Tyr Glu Lys Gln Val Ala Gln Ala Phe Ala Asn Leu Arg Ala Cys 50 55 60Leu Ala Ala Val Gly Ala Thr Ser Asn Asp Ile Thr Lys Leu Asn Tyr65 70 75 80Tyr Ile Val Asp Tyr Asn Pro Ser Lys Leu Thr Ala Ile Gly Asp Gly 85 90 95Leu Lys Ala Thr Phe Ala Leu Asp Arg Leu Pro Pro Cys Thr Leu Val 100 105 110Pro Val Pro Ala Leu Ala Ser Pro Glu Tyr Leu Phe Glu Val Asp Ala 115 120 125Thr Ala Leu Val Pro Gly His Ser Thr Pro Asp Asn Val Ala Asp Val 130 135 140Val Val Val Gly Ala Gly Leu Ser Gly Leu Glu Thr Ala Arg Lys Val145 150 155 160Gln Ala Ala Gly Leu Ser Cys Leu Val Leu Glu Ala Met Asp Arg Val 165 170 175Gly Gly Lys Thr Leu Ser Val Gln Ser Gly Pro Gly Arg Thr Thr Ile 180 185 190Asn Asp Leu Gly Ala Ala Trp Ile Asn Asp Ser Asn Gln Ser Glu Val 195 200 205Phe Lys Leu Phe Glu Arg Phe His Leu Glu Gly Glu Leu Gln Arg Thr 210 215 220Thr Gly Asn Ser Ile His Gln Ala Gln Asp Gly Thr Thr Thr Thr Ala225 230 235 240Pro Tyr Gly Asp Ser Leu Leu Ser Glu Glu Val Ala Ser Ala Leu Ala 245 250 255Glu Leu Leu Pro Ala Trp Ser Gln Leu Ile Glu Glu His Ser Leu Glu 260 265 270Asp Pro Lys Ala Ser Pro Gln Ala Lys Gln Leu Asp Ser Val Ser Phe 275 280 285Ala His Tyr Cys Glu Lys Asp Leu Asn Leu Pro Ala Val Leu Gly Val 290 295 300Ala Asn Gln Ile Thr Arg Ala Leu Leu Gly Val Glu Ala His Glu Ile305 310 315 320Ser Met Phe Phe Leu Thr Asp Tyr Ile Lys Ser Ala Thr Gly Leu Ser 325 330 335Asn Ile Val Ser Asp Lys Lys Asp Gly Gly Gln Tyr Met Arg Cys Lys 340 345 350Thr Gly Met Gln Ser Leu Cys His Ala Met Ser Lys Glu Leu Val Pro 355 360 365Gly Ser Val His Leu Asn Thr Pro Val Ala Glu Ile Glu Gln Ser Ala 370 375 380Ser Gly Cys Thr Val Arg Ser Ala Ser Gly Gly Val Phe Arg Ser Lys385 390 395 400Lys Val Val Val Ser Leu Pro Thr Thr Leu Tyr Pro Thr Leu Ile Phe 405 410 415Ser Pro Pro Leu Pro Ala Glu Lys Gln Ala Leu Ala Glu Lys Ser Ile 420 425 430Leu Gly Tyr Tyr Ser Lys Ile Val Phe Val Trp Asp Lys Pro Trp Trp 435 440 445Arg Glu Gln Gly Phe Ser Gly Val Leu Gln Ser Ser Cys Asp Pro Ile 450 455 460Ser Phe Ala Arg Asp Thr Ser Ile Glu Val Asp Arg Gln Trp Ser Ile465 470 475 480Thr Cys Phe Met Val Gly Asp Pro Gly Arg Lys Trp Ser Gln Gln Ser 485 490 495Lys Gln Val Arg Gln Lys Ser Val Trp Asn Gln Leu Arg Ala Ala Tyr 500 505 510Glu Asn Ala Gly Ala Gln Val Pro Glu Pro Ala Asn Val Leu Glu Ile 515 520 525Glu Trp Ser Lys Gln Gln Tyr Phe Gln Gly Ala Pro Ser Ala Val Tyr 530 535 540Gly Leu Asn Cys Leu Asn Thr Leu Gly Ser Ala Leu Arg Thr Pro Phe545 550 555 560Lys Gly Val His Phe Val Gly Thr Glu Thr Ser Leu Val Trp Lys Gly 565 570 575Tyr Met Glu Gly Ala Ile Arg Ser Gly Gln Arg Gly Ala Ala Glu Val 580 585 590Val Ala Ser Leu Val Pro Ala Ala 595 600 32 1803 DNA Rhinocladiella atrovirens CDS (1)...(1803) misc_feature (0)...(0) isolate C4 32atg gca ctt gca ccg agc tac atc aat ccc cca aac ctc gcc tcc cca 48Met Ala Leu Ala Pro Ser Tyr Ile Asn Pro Pro Asn Leu Ala Ser Pro 1 5 10 15gca ggg tat tcc cac gtc ggc gta ggc cca aac gga ggg agg tat gtg 96Ala Gly Tyr Ser His Val Gly Val Gly Pro Asn Gly Gly Arg Tyr Val 20 25 30aca ata gct gga cag att gga caa gac gct tcg gcc gtg aca gac cct 144Thr Ile Ala Gly Gln Ile Gly Gln Asp Ala Ser Ala Val Thr Asp Pro 35 40 45gcc tac gag aaa cag gtt gcc caa gca ttc gcc aac ctg cga gct tgt 192Ala Tyr Glu Lys Gln Val Ala Gln Ala Phe Ala Asn Leu Arg Ala Cys 50 55 60ctt gct gca gtt gga gcc act tca aac gac att acc aag ctc aat tac 240Leu Ala Ala Val Gly Ala Thr Ser Asn Asp Ile Thr Lys Leu Asn Tyr 65 70 75 80tac atc gtc gac tac aac ccg agc aaa ctc acc gca att gga gat ggg 288Tyr Ile Val Asp Tyr Asn Pro Ser Lys Leu Thr Ala Ile Gly Asp Gly 85 90 95ctg aag gct acc ttt gcc ctt gac agg ctc cct cct tgc acg ctg gtg 336Leu Lys Ala Thr Phe Ala Leu Asp Arg Leu Pro Pro Cys Thr Leu Val 100 105 110cca gtg ccg gcc ctg gct tca cct gaa tac ctc ttt gag gtt gat gct 384Pro Val Pro Ala Leu Ala Ser Pro Glu Tyr Leu Phe Glu Val Asp Ala 115 120 125acg gcg ctg gtt cca gga cac tca acc cca gac aat gtt gcg gac gtg 432Thr Ala Leu Val Pro Gly His Ser Thr Pro Asp Asn Val Ala Asp Val 130 135 140gtc gtg gtg ggc gct ggc ttg agc ggt ttg gag acg gca cgc aaa gtc 480Val Val Val Gly Ala Gly Leu Ser Gly Leu Glu Thr Ala Arg Lys Val145 150 155 160cag gct gcc ggg ctg tcc tgc ctc gtt ctt gag gcg atg gat cgt gtg 528Gln Ala Ala Gly Leu Ser Cys Leu Val Leu Glu Ala Met Asp Arg Val 165 170 175ggg gga aag act ctg agc gta caa tcg ggt ccc ggc agg acg act atc 576Gly Gly Lys Thr Leu Ser Val Gln Ser Gly Pro Gly Arg Thr Thr Ile 180 185 190aat gac ctc ggc gct gcg tgg atc aat gac agc aac caa agc gaa gta 624Asn Asp Leu Gly Ala Ala Trp Ile Asn Asp Ser Asn Gln Ser Glu Val 195 200 205ttc aaa tta ttt gaa aga ttt cat ttg gag ggc gag ctc cag agg acg 672Phe Lys Leu Phe Glu Arg Phe His Leu Glu Gly Glu Leu Gln Arg Thr 210 215 220acc gga aat tca atc cat caa gca caa gac ggt aca acc act aca gct 720Thr Gly Asn Ser Ile His Gln Ala Gln Asp Gly Thr Thr Thr Thr Ala225 230 235 240cct tat ggt gat tcc ctg ctg agc gag gag gtt gca agt gca ctc gcg 768Pro Tyr Gly Asp Ser Leu Leu Ser Glu Glu Val Ala Ser Ala Leu Ala 245 250 255gaa ctc ctt ccc gca tgg tct cag ctg atc gaa gag cat agt ctt gaa 816Glu Leu Leu Pro Ala Trp Ser Gln Leu Ile Glu Glu His Ser Leu Glu 260 265 270gac ccc aag gcg agc cct caa gcg aag cag ctc gac agt gtg agc ttc 864Asp Pro Lys Ala Ser Pro Gln Ala Lys Gln Leu Asp Ser Val Ser Phe 275 280 285gca cac tac tgt gag aag gat cta aac ttg cct gct gtt ctc ggc gtg 912Ala His Tyr Cys Glu Lys Asp Leu Asn Leu Pro Ala Val Leu Gly Val 290 295 300gca aac cag atc aca cgc gct ctg ctc ggt gtg gaa gcc cac gag atc 960Ala Asn Gln Ile Thr Arg Ala Leu Leu Gly Val Glu Ala His Glu Ile305 310 315 320agc atg ctt ttt ctc acc gac tac atc aag agt gcc acc ggt ctc agt 1008Ser Met Leu Phe Leu Thr Asp Tyr Ile Lys Ser Ala Thr Gly Leu Ser 325 330 335aat att gtc tcg gat aag aaa gac ggt ggg cag tat atg cga tgc aaa 1056Asn Ile Val Ser Asp Lys Lys Asp Gly Gly Gln Tyr Met Arg Cys Lys 340 345 350aca ggt atg cag tcg ctt tgc cat gcc atg tca aag gaa ctt gtt cca 1104Thr Gly Met Gln Ser Leu Cys His Ala Met Ser Lys Glu Leu Val Pro 355 360 365ggc tca gtg cac ctc aac acc ccc gtc gcc gaa att gag cag tcg gca 1152Gly Ser Val His Leu Asn Thr Pro Val Ala Glu Ile Glu Gln Ser Ala 370 375 380tcc ggc tgt aca gta cga tcg gcc tcg ggc ggc gtg ttc cga agt aaa 1200Ser Gly Cys Thr Val Arg Ser Ala Ser Gly Gly Val Phe Arg Ser Lys385 390 395 400aag gtg gtg gtt tcg tta ccg aca acc ttg tat ccc acc ttg ata ttt 1248Lys Val Val Val Ser Leu Pro Thr Thr Leu Tyr Pro Thr Leu Ile Phe 405 410 415tca cca cct ctt ccc gcc gag aag caa gca ttg gct gaa aaa tcc atc 1296Ser Pro Pro Leu Pro Ala Glu Lys Gln Ala Leu Ala Glu Lys Ser Ile 420 425 430ctg ggc tac tat agc aag ata gtc ttc gta tgg gac aag ctg tgg tgg 1344Leu Gly Tyr Tyr Ser Lys Ile Val Phe Val Trp Asp Lys Leu Trp Trp 435 440 445cgc gaa caa ggc ttc tcg ggc gtc ctc caa tcg agc tgt gac ccc atc 1392Arg Glu Gln Gly Phe Ser Gly Val Leu Gln Ser Ser Cys Asp Pro Ile 450 455 460tca ttt gcc aga gat acc agc atc gaa gtc gat cgg caa tgg tcc att 1440Ser Phe Ala Arg Asp Thr Ser Ile Glu Val Asp Arg Gln Trp Ser Ile465 470 475 480acc tgt ttc atg gtc gga gac ccg gga cgg aag tgg tcc caa cag tcc 1488Thr Cys Phe Met Val Gly Asp Pro Gly Arg Lys Trp Ser Gln Gln Ser 485 490 495aag cag gta cga cag aag tct gtc tgg aac caa ctc cgc gca gcc tac 1536Lys Gln Val Arg Gln Lys Ser Val Trp Asn Gln Leu Arg Ala Ala Tyr 500 505 510gag aac gcc ggg gcc caa gtc cca gag ccg gcc aac gtg ctc gag atc 1584Glu Asn Ala Gly Ala Gln Val Pro Glu Pro Ala Asn Val Leu Glu Ile 515 520 525gag tgg tcg aag cag cag tat ttc caa gga gcg ccg agc gcc gtc tat 1632Glu Trp Ser Lys Gln Gln Tyr Phe Gln Gly Ala Pro Ser Ala Val Tyr 530 535 540ggg ctg aac tgt ctc aac aca ctg ggt tcg gcg ctc aga acg ccg ttc 1680Gly Leu Asn Cys Leu Asn Thr Leu Gly Ser Ala Leu Arg Thr Pro Phe545 550 555 560aag ggt gtt cat ttc gtt gga acg gag acg tct ttg gtt tgg aaa ggg 1728Lys Gly Val His Phe Val Gly Thr Glu Thr Ser Leu Val Trp Lys Gly 565 570 575tat atg gaa ggg gcc ata cga tcg ggt cag cga ggc gct gca gaa gtt 1776Tyr Met Glu Gly Ala Ile Arg Ser Gly Gln Arg Gly Ala Ala Glu Val 580 585 590gtg cct agc ctg gtg cca gca gca tag 1803Val Pro Ser Leu Val Pro Ala Ala * 595 600 33 600 PRT Rhinocladiella atrovirens 33Met Ala Leu Ala Pro Ser Tyr Ile Asn Pro Pro Asn Leu Ala Ser Pro 1 5 10 15Ala Gly Tyr Ser His Val Gly Val Gly Pro Asn Gly Gly Arg Tyr Val 20 25 30Thr Ile Ala Gly Gln Ile Gly Gln Asp Ala Ser Ala Val Thr Asp Pro 35 40 45Ala Tyr Glu Lys Gln Val Ala Gln Ala Phe Ala Asn Leu Arg Ala Cys 50 55 60Leu Ala Ala Val Gly Ala Thr Ser Asn Asp Ile Thr Lys Leu Asn Tyr65 70 75 80Tyr Ile Val Asp Tyr Asn Pro Ser Lys Leu Thr Ala Ile Gly Asp Gly 85 90 95Leu Lys Ala Thr Phe Ala Leu Asp Arg Leu Pro Pro Cys Thr Leu Val 100 105 110Pro Val Pro Ala Leu Ala Ser Pro Glu Tyr Leu Phe Glu Val Asp Ala 115 120 125Thr Ala Leu Val Pro Gly His Ser Thr Pro Asp Asn Val Ala Asp Val 130 135 140Val Val Val Gly Ala Gly Leu Ser Gly Leu Glu Thr Ala Arg Lys Val145 150 155 160Gln Ala Ala Gly Leu Ser Cys Leu Val Leu Glu Ala Met Asp Arg Val 165 170 175Gly Gly Lys Thr Leu Ser Val Gln Ser Gly Pro Gly Arg Thr Thr Ile 180 185 190Asn Asp Leu Gly Ala Ala Trp Ile Asn Asp Ser Asn Gln Ser Glu Val 195 200 205Phe Lys Leu Phe Glu Arg Phe His Leu Glu Gly Glu Leu Gln Arg Thr 210 215 220Thr Gly Asn Ser Ile His Gln Ala Gln Asp Gly Thr Thr Thr Thr Ala225 230 235 240Pro Tyr Gly Asp Ser Leu Leu Ser Glu Glu Val Ala Ser Ala Leu Ala 245 250 255Glu Leu Leu Pro Ala Trp Ser Gln Leu Ile Glu Glu His Ser Leu Glu 260 265 270Asp Pro Lys Ala Ser Pro Gln Ala Lys Gln Leu Asp Ser Val Ser Phe 275 280 285Ala His Tyr Cys Glu Lys Asp Leu Asn Leu Pro Ala Val Leu Gly Val 290 295 300Ala Asn Gln Ile Thr Arg Ala Leu Leu Gly Val Glu Ala His Glu Ile305 310 315 320Ser Met Leu Phe Leu Thr Asp Tyr Ile Lys Ser Ala Thr Gly Leu Ser 325 330 335Asn Ile Val Ser Asp Lys Lys Asp Gly Gly Gln Tyr Met Arg Cys Lys 340 345 350Thr Gly Met Gln Ser Leu Cys His Ala Met Ser Lys Glu Leu Val Pro 355 360 365Gly Ser Val His Leu Asn Thr Pro Val Ala Glu Ile Glu Gln Ser Ala 370 375 380Ser Gly Cys Thr Val Arg Ser Ala Ser Gly Gly Val Phe Arg Ser Lys385 390 395 400Lys Val Val Val Ser Leu Pro Thr Thr Leu Tyr Pro Thr Leu Ile Phe 405 410 415Ser Pro Pro Leu Pro Ala Glu Lys Gln Ala Leu Ala Glu Lys Ser Ile 420 425 430Leu Gly Tyr Tyr Ser Lys Ile Val Phe Val Trp Asp Lys Leu Trp Trp 435 440 445Arg Glu Gln Gly Phe Ser Gly Val Leu Gln Ser Ser Cys Asp Pro Ile 450 455 460Ser Phe Ala Arg Asp Thr Ser Ile Glu Val Asp Arg Gln Trp Ser Ile465 470 475 480Thr Cys Phe Met Val Gly Asp Pro Gly Arg Lys Trp Ser Gln Gln Ser 485 490 495Lys Gln Val Arg Gln Lys Ser Val Trp Asn Gln Leu Arg Ala Ala Tyr 500 505 510Glu Asn Ala Gly Ala Gln Val Pro Glu Pro Ala Asn Val Leu Glu Ile 515 520 525Glu Trp Ser Lys Gln Gln Tyr Phe Gln Gly Ala Pro Ser Ala Val Tyr 530 535 540Gly Leu Asn Cys Leu Asn Thr Leu Gly Ser Ala Leu Arg Thr Pro Phe545 550 555 560Lys Gly Val His Phe Val Gly Thr Glu Thr Ser Leu Val Trp Lys Gly 565 570 575Tyr Met Glu Gly Ala Ile Arg Ser Gly Gln Arg Gly Ala Ala Glu Val 580 585 590Val Pro Ser Leu Val Pro Ala Ala 595 600

Claims

1. A method of reducing pathogenicity to a plant of a fungus that produces fumonisin, comprising:a) stably integrating into the genome of a plant cell a first nucleotide sequence operably linked to a promoter active in said plant cell, wherein said first nucleotide sequence has at least 95% sequence identity to the sequence set forth in SEQ ID NO: 16, 18, 20, 22, 24, 26, 28, 30, or 32 and encodes a polypeptide having amine oxidase activity; b) optionally stably integrating into the genome of said plant cell a second nucleotide sequence operably linked to a promoter active in said plant cell, wherein said second nucleotide sequence has at least 95% sequence identity to the sequence set forth in SEQ ID NO: 12 or 14 and encodes a polypeptide having fumonisin esterase activity; c) stably integrating into the genome of said plant cell a nucleotide sequence operably linked to a promoter active in said plant cell, wherein said nucleotide sequence has at least 95% sequence identity to the sequence set forth in SEQ ID NO: 2, 4, 7, or 10, and encodes a polypeptide having fumonisin detoxification activity; and d) regenerating a transformed plant from said plant cell, whereby the pathogenicity of said fungus to said transformed plant is reduced in comparison to the pathogenicity of said fungus to a plant that has not been transformed.

2. The method of claim 1, wherein said second nucleotide sequence comprises the sequence set forth in SEQ ID NO: 12 or 14.

3. The method of claim 1, wherein the nucleotide sequence of step (c) comprises the sequence set forth in SEQ ID NO: 2, 4, 7, or 10.

4. The method of claim 1, wherein said first nucleotide sequence comprises the sequence set forth in SEQ ID NO: 16, 18, 20, 22, 24, 26, 28, 30, or 32.

5. The method of claim 1, wherein said plant cell is a cell from a monocot.

6. The method of claim 5, wherein said monocot is maize.

7. The method of claim 1, wherein said plant cell is a cell from a dicot.

8. The method of claim 1, wherein the promoter of step (a) is an inducible promoter.

9. The method of claim 1, wherein the promoter of step (a) is a tissue-preferred promoter.

10. A plant having stably integrated into its genome:a) a first nucleotide sequence operably linked to a promoter active in said plant, wherein said first nucleotide sequence has at least 95% sequence identity to the sequence set forth in SEQ ID NO: 16, 18, 20, 22, 24, 26, 28, 30, or 32 and encodes a polypeptide having amine oxidase activity; b) optionally, a second nucleotide sequence operably linked to a promoter active in said plant, wherein said second nucleotide sequence has at least 95% sequence identity to the sequence set forth in SEQ ID NO: 12 or 14 and encodes a polypeptide having fumonisin esterase activity; and, c) a nucleotide sequence operably linked to a promoter active in said plant, wherein said nucleotide sequence has at least 95% identity to the sequence set forth in SEQ ID NO: 2, 4, 7, or 10, and encodes a polypeptide having fumonisin detoxification activity.

11. The plant of claim 10, wherein said second nucleotide sequence is set forth in SEQ ID NO: 12 or 14.

12. The plant of claim 10, wherein said first nucleotide sequence encodes a polypeptide having the sequence set forth in SEQ ID NO: 17, 19, 21, 23, 25, 27, 29, 31, or 33.

13. The plant of claim 10, wherein said first nucleotide sequence comprises the sequence set forth in SEQ ID NO: 16, 18, 20, 22, 24, 26, 28, 30, or 32.

14. The plant of claim 10, wherein said plant is a monocot.

15. The plant of claim 14, wherein said monocot is maize.

16. The plant of claim 10, wherein said plant is a dicot.

17. Transformed seed of the plant of claim 10.

18. A plant cell having stably integrated into its genome:a) a first nucleotide sequence operably linked to a promoter active in said plant cell, wherein said first nucleotide sequence has at least 95% sequence identity to the sequence set forth in SEQ ID NO: 16, 18, 20, 22, 24, 26, 28, 30, or 32 and encodes a polypeptide having amine oxidase activity; b) optionally, a second nucleotide sequence operably linked to a promoter active in said plant cell, wherein said second nucleotide sequence has at least 95% sequence identity to the sequence set forth in SEQ ID NO: 12 or 14 and encodes a polypeptide having fumonisin esterase activity; and, c) a nucleotide sequence operably linked to a promoter active in said plant cell, wherein said nucleotide sequence has at least 95% identity to the sequence set forth in SEQ ID NO: 2, 4, 7, or 10 and encodes a polypeptide having fumonisin detoxification activity.

19. A method of reducing pathogenicity to a plant of a fungus that produces fumonisin, comprising stably integrating into the genome of a plant cell:a) a first nucleotide sequence operably linked to a promoter active in said plant cell, wherein said first nucleotide sequence encodes a polypeptide having at least 95% sequence identity to the sequence set forth in SEQ ID NO: 17, 19, 21, 23, 25, 27, 29, 31, or 33 and having amine oxidase activity; b) optionally, a second nucleotide sequence operably linked to a promoter active in said plant cell, wherein said second nucleotide sequence encodes a polypeptide having at least 95% sequence identity to the sequence set forth in SEQ ID NO: 13 or 15 and having fumonisin esterase activity; c) a nucleotide sequence operably linked to a promoter active in said plant cell, wherein said nucleotide sequence encodes a polypeptide having at least 95% sequence identity to the sequence set forth in SEQ ID NO: 3, 5, 8, or 11, and having fumonisin detoxification activity; and d) regenerating a transformed plant from said plant cell, whereby the pathogenicity of said fungus to said transformed plant is reduced in comparison to the pathogenicity of said fungus to a plant that has not been transformed.

20. A plant having stably integrated into its genomea) a first nucleotide sequence operably linked to a promoter active in a plant cell, wherein said first nucleotide sequence encodes a polypeptide having at least 95% sequence identity to the sequence set forth in SEQ ID NO: 17, 19, 21, 23, 25, 27, 29, 31, or 33 and having amine oxidase activity b) optionally, a second nucleotide sequence operably linked to a promoter active in a plant cell, wherein said second nucleotide sequence encodes a polypeptide having at least 95% sequence identity to the sequence set forth in SEQ ID NO: 13 or 15 and having fumonisin esterase activity; and, c) a nucleotide sequence operably linked to a promoter active in a plant cell, wherein said nucleotide sequence encodes a polypeptide having at least 95% sequence identity to the sequence set forth in SEQ ID NO: 3, 5, 8, or 11 and having fumonisin detoxification activity.

21. The method of claim 1, wherein the nucleotide sequence of step (c) has at least 98% identity to the sequence set forth in SEQ ID NO: 2, 4, 7, or 10.

22. The method of claim 1, wherein the nucleotide sequence of step (c) encodes the polypeptide set forth in SEQ ID NO: 3, 5, 8, or 11.

23. The plant cell of claim 18, wherein said first nucleotide sequence is set forth in SEQ ID NO: 16, 18, 20, 22, 24, 26, 28, 30, or 32.

24. The plant of claim 10, wherein the nucleotide sequence of step (c) encodes the polypeptide set forth in SEQ ID NO: 3, 5, 8, or 11.

25. The plant of claim 10, wherein the nucleotide sequence of step (c) is the sequence set forth in SEQ ID NO: 2, 4, 7, or 10.

26. The plant cell of claim 18, wherein said first nucleotide sequence has at least 98% sequence identity to the sequence set forth in SEQ ID NO: 16, 18, 20, 22, 24, 26, 28, 30, or 32, said second nucleotide sequence has at least 98% sequence identity to the sequence set forth in SEQ ID NO: 12 or 14, and the nucleotide sequence of step (c) has at least 98% identity to the sequence set forth in SEQ ID NO: 2, 4, 7, or 10.

27. The plant cell of claim 18, wherein the nucleotide sequence of step (c) encodes the polypeptide set forth in SEQ ID NO: 3, 5, 8, or 11.

28. The plant cell of claim 18, wherein said first nucleotide sequence encodes a polypeptide having at least 95% sequence identity to the sequence set forth in SEQ ID NO: 17, 19, 21, 23, 25, 27, 29, 31, or 33, said second nucleotide sequence encodes a polypeptide having at least 95% sequence identity to the sequence set forth in SEQ ID NO: 13 or 15, and the nucleotide sequence of step (c) encodes a polypeptide having at least 95% sequence identity to the sequence set forth in SEQ ID NO: 3, 5, 8, or 11.

29. The method of claim 1, wherein said first nucleotide sequence encodes a polypeptide having the sequence set forth in SEQ ID NO: 17, 19, 21, 23, 25, 27, 29, 31, or 33, said second nucleotide sequence encodes a polypeptide having the sequence set forth in SEQ ID NO: 13 or 15, and the nucleotide sequence of step (c) encodes a polypeptide having the sequence set forth in SEQ ID NO: 3, 5, 8, or 11.

30. The plant cell of claim 18, wherein said first nucleotide sequence encodes a polypeptide comprising the sequence set forth in SEQ ID NO: 17, 19, 21, 23, 25, 27, 29, 31, or 33, said second nucleotide sequence encodes a polypeptide comprising the sequence set forth in SEQ ID NO: 13 or 15, and the nucleotide sequence of step (c) encodes a polypeptide comprising the sequence set forth in SEQ ID NO: 3, 5, 8, or 11.

31. The plant cell of claim 18, wherein the nucleotide sequence of step (c) is set forth in SEQ ID NO: 2,4, 7, or 10.

32. The plant of claim 10, wherein said first nucleotide sequence encodes a polypeptide comprising the sequence set forth in SEQ ID NO: 17, 19, 21, 23, 25, 27, 29, 31, or 33, said second nucleotide sequence encodes a polypeptide comprising the sequence set forth in SEQ ID NO: 13 or 15, and the nucleotide sequence of step (c) encodes a polypeptide comprising the sequence set forth in SEQ ID NO: 3, 5, 8, or 11.

33. The plant cell of claim 18, wherein said second nucleotide sequence is set forth in SEQ ID NO: 12 or 14.