CN116286897B - Tri3 gene for encoding acyltransferase and application thereof - Google Patents
Tri3 gene for encoding acyltransferase and application thereof Download PDFInfo
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- CN116286897B CN116286897B CN202310323328.6A CN202310323328A CN116286897B CN 116286897 B CN116286897 B CN 116286897B CN 202310323328 A CN202310323328 A CN 202310323328A CN 116286897 B CN116286897 B CN 116286897B
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/1025—Acyltransferases (2.3)
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- C—CHEMISTRY; METALLURGY
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- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
- C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
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- C12R2001/00—Microorganisms ; Processes using microorganisms
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- C12R2001/85—Saccharomyces
- C12R2001/865—Saccharomyces cerevisiae
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- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The invention relates to a Tri3 gene for encoding acyltransferase and application thereof, and relates to the field of genetic engineering. The nucleotide sequence of the Tri3 gene is shown as SEQ ID NO:1, the acyltransferase is useful for antifungal toxins. The Tri3 gene is useful against mycotoxins. The method lays a molecular biology foundation for improving the capability of saccharomyces cerevisiae anti-gliotoxin and derivatives and trichothecene toxoids in the later period, improving the heterologous expression level of related mycotoxins and analyzing the action mechanism of the related mycotoxins.
Description
Technical Field
The invention relates to the field of genetic engineering, in particular to a Tri3 gene for encoding acyltransferase and application thereof.
Background
Mycotoxins are a class of secondary metabolites produced by toxigenic fungi during growth, and mainly include aflatoxins, deoxynivalenol (vomitoxin), zearalenone, fumonisins, ochratoxins, and the like. How to safely and reasonably utilize the grain crops polluted by toxins and maintain the quality safety of the national grain foods is a problem to be solved urgently. In particular, trichothecene toxins among mycotoxins, which are considered to be the most dangerous naturally occurring food pollutants, have become a hotspot in international research. Trichothecene toxins are a general term for a large class of toxic substances produced by fusarium that have similar chemical structures, and their basic structures are tetracyclic sesquiterpenes, which can be classified into A, B, C, D types according to the substituents. Approximately 170 trichothecenes have been found so far, while the highest contamination rates of trichothecenes type a and B, including DON, 3-ADON, 15-ADON, NIV, T-2, DAS toxins, etc., are the highest contamination rates and levels of DON, and thus, it is a main study content to prevent and reduce the entry of DON toxins into the food chain of humans and animals to prevent their hazard.
Meanwhile, gliotoxin (gliotoxin, GT) is a diketopiperazine compound (epipolythiodioxopiperazine, ETP), ETP is an important virulence factor, and specific toxicity is generated on various cells through different routes, so that important synergistic effect is exerted in invasive aspergillosis, and the gliotoxin is also a mycotoxin of great concern. ETP exerts toxic and side effects, mainly through disulfide bonds, cross-linking of sulfhydryl groups with target proteins, thereby inactivating the activity of the proteins, and can generate toxic active oxygen (reactiveoxygenspecies, ROS) through redox cycling, and the ROS generation mechanism is considered as a mechanism for GT cytotoxicity. The toxicity of gliotoxin to the host mainly includes: inducing apoptosis; resulting in an imbalance in the redox reaction; inhibiting proteasome activity, inhibiting NF- κB activity, and reducing immunity. It is not only toxic to animal and plant cells, but also to host bacteria.
Disclosure of Invention
In view of the above problems, the present invention provides a Tri3 gene encoding an acyltransferase, which Tri3 gene is useful against mycotoxins.
In order to achieve the above object, the present invention provides a Tri3 gene encoding an acyltransferase, wherein the nucleotide sequence of the Tri3 gene is as shown in SEQ ID no: 1, the acyltransferase is useful for antifungal toxins.
The invention also provides an acyltransferase for antifungal toxins, which is encoded by the Tri3 gene.
The invention also provides an expression vector containing the Tri3 gene.
In one embodiment, the expression vector is a gene expression cassette.
In one embodiment, the initial vector of the expression vector is a YEp352-TEF1-CYC1 plasmid vector.
The invention also provides an expression strain, which contains the expression vector.
In one embodiment, the starting strain of the expression strain is Saccharomyces cerevisiae BJ5464.
The invention also provides application of the Tri3 gene in antifungal toxin.
The invention also provides application of the Tri3 gene in preparing antifungal toxin products.
The invention also provides application of the acyltransferase in antifungal toxin.
The invention also provides application of the acyltransferase in preparing antifungal toxin products.
In one embodiment, the mycotoxin is a gliotoxin, a gliotoxin derivative, or a trichothecene toxin.
In one embodiment, the trichothecene toxins include at least 1 of vomitoxin, zearalenone, and cephalosporin.
In one embodiment, the cephalosporin is cephalosporin E.
Compared with the prior art, the invention has the following beneficial effects:
The invention relates to a Tri3 gene for encoding an acyltransferase and application thereof, wherein the Tri3 gene can be used for resisting mycotoxins. The method lays a molecular biology foundation for improving the capability of saccharomyces cerevisiae anti-gliotoxin and derivatives and trichothecene toxoids in the later period, improving the heterologous expression level of related mycotoxins and analyzing the action mechanism of the related mycotoxins.
Drawings
FIG. 1 is an electrophoresis chart of the amplification product of the gene Tri3 using the A553 cDNA library as a template in example 1;
FIG. 2 is a map of the YEp352-TEF1-CYC1 vector of example 2;
FIG. 3 is a map of the YEp352-TEF1-Tri3 vector in example 2;
FIG. 4 is an electrophoretogram of PCR amplification products of colonies of Saccharomyces cerevisiae BJ5464-D cells containing the YEp352-TEF1-Tri3 plasmid of example 2;
FIG. 5 is a graph showing the effect of Saccharomyces cerevisiae BJ5464-D (YEp 352-TEF1-CYC 1), saccharomyces cerevisiae BJ5464-D (YEp 352-TEF1-Tri 3) in example 2 cultured for 30 hours in YPD plates without any toxin added, and YPD plates containing mycotoxins (including 2.5. Mu.M FS140-12-2, 250. Mu.M DON, 250. Mu.M ZEN, and 200. Mu.M MR 4), wherein the plates are in this order from left to right: YPD plates without any toxin added, YPD plates with 2.5. Mu.M FS140-12-2, YPD plates with 250. Mu.M DON, YPD plates with 250. Mu.M ZEN, YPD plates with 200. Mu.M MR4, A in each plate being Saccharomyces cerevisiae BJ5464-D (YEp 352-TEF1-CYC 1) and C being Saccharomyces cerevisiae BJ5464-D (YEp 352-TEF1-Tri 3).
Detailed Description
In order that the invention may be readily understood, a more complete description of the invention will be rendered by reference to the appended drawings. Preferred embodiments of the present invention are shown in the drawings. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.
The reagents, materials and equipment used in the examples are all commercially available sources unless otherwise specified; the experimental methods are all routine experimental methods in the field unless specified.
Example 1
Acquisition of Tri3 Gene sequence encoding an Acyltransferase.
Endophytic fungi Paramyrothecium roridum A of the invention are disclosed in the following documents and can be obtained according to the literature ：Wei Ye#,Muzi Zhu#,Saini Li#,Youfei Cen,Taomei Liu,Haohua Li,Hongxin Liu,Weimin Zhang*.The excavation of novel toxinresistance proteins against Trichothecenes toxins in Paramyrothecium roridum.International Journal of Biological Macromolecules,2021,192:369–378,https://doi.org/10.1016/j.ijbiomac.2021.09.185.
In this example, the formulation of YPD solid medium is: each liter contains yeast powder 10g, peptone 20g, glucose 20g and agar powder 20g, and the balance is distilled water, and the preparation method is that the components are uniformly mixed and sterilized.
Inoculating the endophytic fungi Paramyrothecium roridum A to a YPD culture medium plate, culturing at 37 ℃ for 72 hours, picking up fresh mycelium, extracting RNA by using a fungus RNA extraction kit, and carrying out reverse transcription by using All-in-one RT MASTER KIT to obtain cDNA. Based on the result of transcriptome sequencing, predicting the sequence of the gene Tri3 encoding the acylase, designing the upstream primer Tri3-F and the downstream primer Tri3-R, wherein the specific sequences are as follows:
Tri3-F:5'-ATGGGTAGCCTGCCCGCATT-3'(SEQ ID NO：2)；
Tri3-R:5'-TCACAGTCTGAAGGCCAGCATGAAG-3'(SEQ ID NO：3)。
then amplified using the cDNA library as a template to obtain a PCR product, the result of which is shown in FIG. 1.
Recovering the product, performing TA cloning by using a pEASY-T1 kit, transforming into escherichia coli competent cells, coating the escherichia coli competent cells on an ampicillin resistance plate to screen out positive clones, performing bacterial liquid PCR (polymerase chain reaction) verification positive clones by using a universal primer M13-F, M-R, and sequencing to obtain a target gene Tri3 gene, wherein the nucleotide sequence of the Tri3 gene is shown as SEQ ID NO:1 is shown in the specification; the sequence of the universal primer is shown in SEQ ID NO: 4. SEQ ID NO: shown at 5.
ATGGGTAGCCTGCCCGCATTCCAACTGCCCCCATTGATCCCCGAGAACCACCGGTGGAACATATCCAAGACGAACCCGCGCCTGGCGCGACGAAGGGGCATCGGCTTCGAGGTCATTGTAGGGTTCGAGCAGTTGAATCGACGGGGCCAGTACGACCTGTACCTCACCGCGTCGCTCCGCACCGTCGACGCTCCAGGATCCACAGCCCTGTCCCTAGCATACCTGAAGGAGAAGTTCGAATCAACCCTGTTGGTGGCGCGCCATGAACATCCCGAGTGTGCTTGCACTGCTCACTGGGATGAGAAGCCCAATCCCATTTTTGAGTACGAGTCGCCCGAGAGCGACGAGGCGGCACTTGCGTGGGCCAAGGACACCGTCCATGCGGTGCCCACCGCCCAGACGGCCCAGCAGGTTTGGTACGATATTGAACGGCGGCGACAGCAGACTGCCGTCGCGGACCGCAAGCCTGGAAAGCCCGTGGACATTTTCTTGATCTCGGACGTGCCAGACGAAAGCGCTCCGCTCCCCGTGGGGGCGTCGGTCGACGTCCTGTTCCACATGAACCACCTATACTGGGACGGCATCGGGGCGCGCATCGCCGTCGGCTACCTGCTCCGCCAGCTGAACAACTTCATCGGAACCCCTGCCGGACAGGCGCGTCCGAAGATCCAGTGGGGTACGGAACTCTCAAACTTTCACACGGCGGCGATGGACGCGATGAAGATCAAGATTGAGACGCTCGGGGCTGAGTTTGAGGCCCGCGGCCAGCAATACGTAAACACATTGGCGCAGCATGTTAGCTCCCGCGGCATGCCGTTCAAGGCCAGCGACGAGGCGATCCCTCGCGCGCACGTCCTGACCTTCACCCCGGACGAGACCAAGGACATCATCCGCGCCGTCAAGACCCGGCTCGGCCCCAAGTTCACCATCTCGCACCTGGCGCAGGCCGCCACGGTCGTCGCGATGCTCGCCACCTACAAGCCCACGACGGAGGTGTCCGACACGGAGTCCTTCGTGGCGCCCACGGCCGTCAACGCCCGCCGGTACCTGCGGGAGGACCTCAAGGACAGCTACATGGCTGGGTGCGTGACGGGTGCCGTTATCAAGGTCGACAACGTCAAGTCGCTGCTCGTCACGCTCGACGACGACAAGGACGCCGTCGTCGCGGCCCTGGCGAGCGCCACCAAGGACGTCAAGGCCTCATTCGACCTGTGGATTCAGGACCCGTCCCAGCTCGCCCTCGGCCTCCGAACCCTCACCTTTGAGGGCATGATGCTGGCCAGAAACCCTATGCCGTTTGACAAGACGTCCGGGCCGTTCATCTCAAGCGACGGCATCAACGAGCAGTACATCCCTACGGCAGTGACGTCGACAGCTACGGGCGAGACTCTTTTGAAGACTGTCAACTTCACCTTCTTGCTCAACCAGTTCCTTCCATATGTTGCACTCCGGCTGGACAGCTGGAACGACACGTCGATGCTGACCATCTGCTACAACGACGCCAATTTTACGGAAGAGGAGACGGCCAAGTATCTCAAGTCCGTTGCGGACTTCATGCTGGCCTTCAGACTG(SEQ ID NO：1);
M13-F：5′-GTAAAACGACGGCCAGT-3′(SEQ ID NO：4)；
M13-R：5′-CAGGAAACAGCTATGAC-3′(SEQ ID NO：5)。
Example 2
Functional verification of the Tri3 gene encoding an acyltransferase.
The Tri3 gene encoding an acyltransferase was inserted into the yeast vector YEp352-TEF1-CYC1 using homologous recombination. The plasmid constructed in the earlier stage research of the yeast vector YEp352-TEF1-CYC1 carries a constitutive promoter TEF1 and a terminator CYC1, the vector is shown in figure 2, disclosed in the following literature, and can be obtained according to the content of the literature ：Xiaodan Ouyang,Yaping Cha,Wen Li,Chaoyi Zhu,Muzi Zhu,Shuang Li,Min Zhuo,Shaobin Huang and Jianjun Li.Stepwise engineering of Saccharomyces cerevisiaeto produce(+)-valencene and its relatedsesquiterpenes,RSC Adv.,2019,9,30171,DOI:10.1039/c9ra05558d.
First, the upstream and downstream primers YEp352-Tri3-F and YEp352-Tri3-R amplified against the gene Tri3 (SEQ ID NO: 1) were designed, the primer sequences of which are shown below:
YEp352-Tri3-F:5'-GCAATCTAATCTAAGTCTAGAATGGGTAGCCTGCCCGCATT-3'(SEQ ID NO：6)；
YEp352-Tri3-R:5'-TACATGATGCGGCCCGTCGACTCACAGTCTGAAGGCCAGCATGAAG-3'(SEQ ID NO：7)。
the underlined sequences of the upstream and downstream primers YEp352-Tri3-F and YEp352-Tri3-R are homologous arm fragments, and the products are obtained through PCR amplification.
The vector YEp352-TEF1-CYC1 was digested with Sal I and Xba I and the products recovered, and then the two products were recombinantly ligated and transformed into DH 5. Alpha. Using ClonExpress II One Step Cloning Kit C (Vazyme) to screen for positive clones. Colony PCR verification was performed using the primers YEp352-Tri3-F and YEp352-Tri3-R, and the results showed that the gene Tri3 was successfully inserted between the promoter TEF1 and the terminator CYC1, into the YEp352-TEF1-CYC1 vector (FIG. 2), and confirmed by sequencing, resulting in the YEp352-TEF1-Tri3 vector (vector map see FIG. 3).
Competent cells of toxin-sensitive Saccharomyces cerevisiae Saccharomyces cerevisiaeBJ5464-D (relevant genotype: Δpdr5Δpdr10Δpdr15) which are more sensitive to toxic compounds and which have been disclosed in the literature, were prepared by electrotransferring ：Wolfgang Schweiger,Jayanand Boddu,Sanghyun Shin,Brigitte Poppenberger,Franz Berthiller,Marc Lemmens,Gary J.Muehlbauer,and Gerhard Adam.Validation of a Candidate Deoxynivalenol-InactivatingUDP-Glucosyltransferase from Barleyby Heterologous Expression in Yeast,MPMI,2010,Vol.23,No.7,DOI:10.1094/MPMI-23-7-0977. a YEp352-TEF1-Tri3 plasmid vector and a YEp352-TEF1-CYC1 plasmid vector (negative control) into Saccharomyces cerevisiae BJ5464-D cells, respectively, (1500V, 5 ms), uniformly plating them on uracil-deficient SD plates, culturing 2D at 30℃and screening positive clones by colony PCR to obtain an electrophoretogram of the PCR amplified products of Saccharomyces cerevisiae BJ5464-D cells containing the YEp352-TEF1-Tri3 plasmid and the YEp352-TEF1-CYC1 plasmid, respectively, as shown in FIG. 4.
Saccharomyces cerevisiae BJ5464-D (YEp 352-TEF1-CYC 1) and Saccharomyces cerevisiae BJ5464-D (YEp 352-TEF1-Tri 3) were inoculated into SD medium, and cultured at 30deg.C for 2D. Each bacterial liquid OD 600 was measured by a spectrophotometer, and each bacterial liquid was diluted with sterile water to OD 600. Apprxeq.1.0 as a stock solution, and diluted to 10 -1 by adding 900. Mu.L of sterile water to 100. Mu.L of stock solution, and diluted to 10 -2、10-3、10-4 in the same manner. mu.L of 10 -2、10-3、10-4 dilutions of each of the different strains were spotted on YPD plates and YPD-various mycotoxin plates, respectively, and incubated at 30℃and observed in real time.
In the present example, the mycotoxins used are Zearalenone (ZEN), vomitoxin (DON), and a derivative of gliotoxin (FS 140-12-2) purchased from Sigma, and the derivative of gliotoxin (FS 140-12-2) was isolated from the deep sea fungus Geosmia PALLIDA FS140, as described in the following, and is obtainable according to the literature ：Zhang-Hua Sun,Jiangyong Gu,Wei Ye,Liang-Xi Wen,Qi-Bin Lin,Sai-Ni Li,Yu-Chan Chen,Hao-Hua Li,Wei-Min Zhang.Geospallins A–C:New Thiodiketopiperazines with Inhibitory Activity against Angiotensin-Converting Enzyme from a Deep-Sea-Derived Fungus Geosmithia pallida FS140.Marine Drugs 2018,16(12),464.https://doi.org/10.3390/md16120464.
In this example, the SD solid medium was formulated as follows: the preparation method comprises the steps of uniformly mixing the components and sterilizing, wherein each liter of the preparation contains 20g of glucose, 0.62g of Do supplement (-Leu/-Trp/-Ura, clontech), 6.7g of non-amino yeast nitrogen source YNB (Praboxin), 0.06g of leucine, 0.04g of tryptophan and 20g of agar powder, and the balance of distilled water.
The plate results of the culture for 30h are shown (FIG. 5), wherein A is Saccharomyces cerevisiae BJ5464-D (YEp 352-TEF1-CYC 1), and C is Saccharomyces cerevisiae BJ5464-D (YEp 352-TEF1-Tri 3); saccharomyces cerevisiae BJ5464-D (YEp 352-TEF1-CYC 1), saccharomyces cerevisiae BJ5464-D (YEp 352-TEF1-Tri 3) grew nearly consistently on YPD plates without any toxin added, but on YPD plates containing mycotoxins (including 2.5. Mu.M FS140-12-2, 250. Mu.M DON, 250. Mu.M ZEN, and 200. Mu.M MR 4), the negative control BJ5464-D (YEp 352-TEF1-CYC 1) grew significantly blocked and was almost never long. The saccharomyces cerevisiae introduced with the novel acylase Tri3 grows well, the thallus density of the saccharomyces cerevisiae at different dilutions is equivalent to that of the normal saccharomyces cerevisiae, and the novel acylase Tri3 partially or completely restores the tolerance of the saccharomyces cerevisiae BJ5464-D to exogenously added toxins, thereby effectively helping the normal growth of the saccharomyces cerevisiae in the environment containing the toxins.
In summary, the present inventors predicted the sequence encoding the acylase Tri3 by the result of transcriptome sequencing, designed specific primers on the upstream and downstream thereof, and obtained the product by PCR amplification using cDNA library obtained by reverse transcription of the transcriptome of endophytic fungus P.roridum A553 as a template and purified and recovered the fragment to obtain the acylase gene Tri3, the nucleotide sequence of which is shown as SEQ ID NO. 1. Then, the Tri3 gene was inserted into the expression cassette of the yeast vector YEp352-TEF1-CYC1 by using the homologous recombination method, which was performed as follows: first, the upstream and downstream primers of the Tri3 gene containing homology arms were designed, and then the products were obtained by PCR amplification and the recovered fragments were purified. The constructed YEp352-TEF1-CYC1 vector is digested with the enzymes Sal I and Xba I, and then the fragments and the digested vector are recombined and connected by ClonExpress II One Step Cloning Kit C (Vazyme) and transformed into competent cells of escherichia coli, and the competent cells are coated on an ampicillin resistance plate to screen positive clones. Through the round of molecular cloning, the target gene Tri3 is inserted between a promoter TEF1 and a terminator CYC1, a YEp352-TEF1-Tri3 vector is constructed, the vector is electrically transferred into a saccharomyces cerevisiae BJ5464-D cell, and uracil-deficient SD culture medium plates are used for screening and verification. Compared with Saccharomyces cerevisiae BJ5464-D transferred with YEp352-TEF1-CYC1 plasmid (negative control), the growth speed of Saccharomyces cerevisiae containing recombinant vector YEp352-TEF1-GliT is obviously accelerated, the colony density is higher in the same culture time, and the functional gene Tri3 is proved to be capable of effectively assisting Saccharomyces cerevisiae to resist exogenous gliotoxin derivatives and trichothecene toxins, so that a foundation is laid for reconstructing gliotoxin and derivatives thereof and trichothecene toxin biosynthesis paths in Saccharomyces cerevisiae.
Endophytic fungi Paramyrothecium roridum A related to the invention are separated from patchouli leaf parts, and the inventor performs transcriptome sequencing on the strain and annotates genes related to trichothecene toxin biosynthesis in earlier researches. The inventor obtains an acyltransferase Tri3 gene sequence from a cDNA library of P.roridum A553 through the operation, and successfully introduces the acyltransferase Tri3 gene sequence into Saccharomyces cerevisiae S.cerevisiae eBJ5464 for antitoxic function verification, thereby providing a new idea for improving the capabilities of Saccharomyces cerevisiae antiglaotoxin and derivatives thereof and trichothecene toxoids in the later period, improving the heterologous expression level of the mycotoxins, analyzing the action mechanism of the mycotoxins, laying a molecular biological foundation, and being beneficial to separating and purifying the novel acyltransferase Tri3 in the next step, determining whether the acyltransferase Tri3 has degradation effect on the mycotoxins, and providing a new idea for developing economic and feasible toxin reducing agents, mycotoxin prevention and control additives and the like.
The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The above examples illustrate only a few embodiments of the invention, which are described in detail and are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.
Claims (10)
1. A Tri3 gene encoding an acyltransferase, wherein the nucleotide sequence of the Tri3 gene is as set forth in SEQ ID NO: 1.
2. An acyltransferase for use in antifungal toxins, wherein the acyltransferase is encoded by the Tri3 gene of claim 1.
3. An expression vector comprising the Tri3 gene of claim 1.
4. The expression vector of claim 3, wherein the initial vector of the expression vector is a YEp352-TEF1-CYC1 plasmid vector.
5. An expression strain comprising the expression vector of any one of claims 3-4.
6. The expression strain according to claim 5, wherein the starting strain of the expression strain is endophytic fungus Paramyrothecium roridum A, or Saccharomyces cerevisiae BJ5464.
7. Use of the Tri3 gene according to claim 1 in antifungal toxins, which are zearalenone, vomitoxin, the gliotoxin derivative FS140-12-2 and/or the cephalosporin E isomer MR4.
8. Use of the Tri3 gene according to claim 1 for the preparation of an antifungal toxin product, said mycotoxin being zearalenone, vomitoxin, a gliotoxin derivative FS140-12-2 and/or a cephalosporin E isomer MR4.
9. Use of an acyltransferase according to claim 2 in an antifungal toxin which is zearalenone, vomitoxin, a gliotoxin derivative FS140-12-2 and/or a cephalosporin E isomer MR4.
10. Use of an acyltransferase according to claim 2 for the preparation of an antifungal product, the mycotoxin being zearalenone, vomitoxin, a gliotoxin derivative FS140-12-2 and/or a cephalosporin E isomer MR4.
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