绿僵菌异戊烯基转移酶基因的克隆及表达谱分析  

Cloning and expression of prenyl transferase gene of Metarhizium ansopliae

作　　者：殷从松[1] 金凯[1] 夏玉先[1] 

机构地区：[1]重庆大学生物工程学院基因工程研究中心,重庆市功能基因及调控技术重点实验室,重庆400030

出　　处：《微生物学报》2010年第7期897-902,共6页Acta Microbiologica Sinica

基　　金：国家自然科学基金(30771446)~~

摘　　要：【目的】克隆绿僵菌异戊烯基转移酶(Mpt)基因,了解该基因的结构和表达特征。【方法】采用SMART(Switching Mechanism At 5′end of RNA Transcript)技术和PCR技术,扩增Mpt基因全长cDNA序列和DNA序列,用qRT-PCR方法分析该基因在绿僵菌不同侵染时期的表达谱。【结果】Mpt基因含2个外显子和1个内含子,CDS区为1026bp(GenBank登录号GU271134),编码341个氨基酸;qRT-PCR分析表明,该基因在绿僵菌侵染的不同时期表达水平有显著差异,特别是在寄主昆虫体内生长的后期高表达。【结论】首次克隆了绿僵菌的Mpt基因,弄清了该基因具有在侵染后期高表达等特征,为研究该基因的功能奠定了基础。[Objective]To clone and identify prenyl transferase gene from Metarhizium anisopliae and to understand the gene structure and expression.[Methods ]Using Switching Mechanism At 5＇ end of RNA Transcript （SMART） method,we isolated the full length cDNA sequence and DNA sequence.Then we used quantitative RT-PCR analysis of the gene expression levels at different stages of colonization of host hemolymph by M.anisopliae.[Results]The Mpt gene had two exons and one intron and the CDS was 1026 bp which encoded a protein with 341 amino acid residues;qRT-PCR analysis showed that the gene expression levels were significantly different,especially highly up-regulated at the late stages.[Conclusion] The Mpt gene was successfully cloned from M.anisopliae for the first time and the gene had the characteristic of high expression levels at the late stages.

关 键 词：昆虫病原真菌 异戊烯基转移酶 SMART技术 QRT-PCR 表达谱 

分 类 号：S476.12[农业科学—农业昆虫与害虫防治] Q93[农业科学—植物保护]