Paramyrothecium roridum中单端孢霉烯毒素生物合成基因启动子的克隆和功能鉴定

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

利用染色体步移技术对植物内生真菌Paramyrothecium roridum中单端孢霉烯毒素的生物合成基因tri5、tri12启动子进行克隆,分别利用原核和真核表达系统以及荧光素酶表达系统对启动子核心区域进行鉴定,发现tri12的启动子片段12-f1对氨苄青霉素抗性基因和潮霉素抗性基因表达的启动效率最高,tri5的启动子片段5-f0对荧光素酶基因表达的启动效率最高;同时对启动子的功能组件进行分析,发现tri5含有TATA box和CAAT box,而tri12是TATA-less启动子,但含有CAAT box和GC box。本研究为强启动子的筛选及单端孢霉烯类化合物的高效生物合成奠定基础。

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.