Putative Nonribosomal Peptide Synthetase and Cytochrome P450 Genes Responsible for Tentoxin Biosynthesis in Alternaria alternata ZJ33
Abstract
:1. Introduction
2. Results
2.1. Identification of Isolate ZJ33
2.2. PCR Amplification of NRPS Gene Fragments
2.3. Characterization of TES and TES1
2.4. Targeted Disruption of TES and TES1 in A. alternata ZJ33
3. Discussion
4. Experimental Section
4.1. Strains, Media, and Culture Conditions
4.2. Nucleic Acid Manipulations
4.3. Identification of Fungi
4.4. Cloning of NRPS Genes from A. alternata ZJ33
4.5. Accession Numbers
4.6. Double-Joint PCR (DJ-PCR) for Constructing Mutant Alleles
4.7. Targeted Mutagenesis
4.8. High-Performance Liquid Chromatography (HPLC) Analysis of Tentoxin
Supplementary Materials
Acknowledgments
Author Contributions
Conflict of Interest
References
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Domain | Position | Substrate | ||||||||
---|---|---|---|---|---|---|---|---|---|---|
235 | 236 | 239 | 278 | 299 | 301 | 322 | 330 | 331 | ||
A1 | D | I | A | Q | V | G | V | I | W | Gly |
A2 | D | V | W | F | C | G | G | T | F | Ala |
A3 | D | A | L | L | V | G | A | V | S | Leu |
A4 | D | G | W | F | L | A | A | V | M | DPhe |
Primer a | Sequence b (5′→3′) |
---|---|
ITS4 | TCCTCCGCTTATTGATATGC |
ITS5 | GGAAGTAAAAGTCGTAACAAGG |
cps1 | AATCTAGATAYGGNCCNACNGA |
cps2 | CCTCTAGANAGRTCNCCNGTYTTR |
NRPS-for | GAGGCAAGGCAACCGCAACGATGA |
NRPS-rev | CCCTTCATGTCGGGACTTGCGACA |
HygB-for | CCGGGCTGCAGGAATTCGAT |
HygB-rev | GGATCCCGGTCGGCATCTAC |
TES-5for | CGGGATCGCTACTGTTTGACGTCA |
TES-5rev | ATCGAATTCCTGCAGCCCGGTTGCCTTGCCTCAGCCAGA |
TES-3for | GTAGATGCCGACCGGGATCCGCTAAAGTGGACCGGCAGA |
TES-3rev | CATCGATCCCGATTGGCGTTCACA |
TES-5nest | GAAGATATGGGAGAGAAACCGCGG |
TES-3nest | AATGGTGGCATCGTTCTGGCCAGT |
P450-5for | ACCTGACGAAACTCACTGCCTCCT |
P450-5rev | ATCGAATTCCTGCAGCCCGGCTCAGGCTCCATCTTTGGAG |
P450-3for | GTAGATGCCGACCGGGATCCAGGGCCTGGAGGAAACCTACAC |
P450-3rev | GCTCGACCATTGGATACCTAGCCT |
P450-5nest | ACAATGGCCTAAGTCTGCCGCTCA |
P450-3nest | CCCTCAACATTCCCTGGCATCTTG |
Hyg-5rev | CGCACAAGTTATCGTGCACCAAGC |
Hyg-3for | GGCGTATATGCTCCGCATTGGTCT |
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Li, Y.-H.; Han, W.-J.; Gui, X.-W.; Wei, T.; Tang, S.-Y.; Jin, J.-M. Putative Nonribosomal Peptide Synthetase and Cytochrome P450 Genes Responsible for Tentoxin Biosynthesis in Alternaria alternata ZJ33. Toxins 2016, 8, 234. https://doi.org/10.3390/toxins8080234
Li Y-H, Han W-J, Gui X-W, Wei T, Tang S-Y, Jin J-M. Putative Nonribosomal Peptide Synthetase and Cytochrome P450 Genes Responsible for Tentoxin Biosynthesis in Alternaria alternata ZJ33. Toxins. 2016; 8(8):234. https://doi.org/10.3390/toxins8080234
Chicago/Turabian StyleLi, You-Hai, Wen-Jin Han, Xi-Wu Gui, Tao Wei, Shuang-Yan Tang, and Jian-Ming Jin. 2016. "Putative Nonribosomal Peptide Synthetase and Cytochrome P450 Genes Responsible for Tentoxin Biosynthesis in Alternaria alternata ZJ33" Toxins 8, no. 8: 234. https://doi.org/10.3390/toxins8080234
APA StyleLi, Y.-H., Han, W.-J., Gui, X.-W., Wei, T., Tang, S.-Y., & Jin, J.-M. (2016). Putative Nonribosomal Peptide Synthetase and Cytochrome P450 Genes Responsible for Tentoxin Biosynthesis in Alternaria alternata ZJ33. Toxins, 8(8), 234. https://doi.org/10.3390/toxins8080234