Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (8): 85-94.doi: 10.13560/j.cnki.biotech.bull.1985.2020-1386

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Cloning and Functional Identification of Trichothecene Mycotoxin Biosynthesis Gene Promoter from Paramyrothecium roridum

CEN You-fei1,2(), ZHU Mu-zi2, YE Wei2, LI Sai-ni2, ZHONG Guo-hua1(), ZHANG Wei-min2()   

  1. 1. Ministry of Education Key Laboratory of Natural Pesticide and Chemical Biology,Ministry of Agriculture and Rural Affairs Key Laboratory of Crop Integrated Pest Management in South China,South China Agricultural University,Guangzhou 510642
    2. State Key Laboratory of Applied Microbiology in Southern China,Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application,Guangdong Open Laboratory of Applied Microbiology,Guangdong Institute of Microbiology,Guangdong Academy of Sciences,Guangzhou 510070
  • Received:2020-11-14 Online:2021-08-26 Published:2021-09-10
  • Contact: ZHONG Guo-hua,ZHANG Wei-min E-mail:297288514@qq.com;guohuazhong@scau.edu.cn;wmzhang@gdim.cn

Abstract:

The promoters of trichothecene mycotoxin biosynthetic genes tri5 and tri12 from endophytic fungus Paramyrothecium roridum were cloned by genome walking,and the core regions of the promoters were identified using prokaryotic,eukaryotic and luciferase expression systems,respectively. The results showed that 12-f1,the promoter fragment of tri12,was of the highest transcriptional efficiency for the expression of ampicillin resistance genes and hygromycin resistance genes;meanwhile,5-f0,the promoter fragment of tri5,demonstrated the highest transcriptional efficiency for luciferase gene expression. Analysis of the functional components of the promoters revealed that tri5 contained TATA box and CAAT box,while tri12 was a TATA-less promoter,but contained CAAT box and GC box. This study lays a foundation for the screening of strong promoters and efficient biosynthesis of trichothecenes.

Key words: Paramyrothecium roridum, trichothecene mycotoxin, promoter, cloning, identification