粉棒束孢类枯草杆菌蛋白酶结构基因及其上游序列的克隆与分析  

Cloning and sequence analysis of structural gene and upstream sequence of subtilisin-like protease from Isaria farinosa

作　　者：庄宗兰[1] 谢翎[2] 马忠友[1] 黄勃[1] 

机构地区：[1]安徽农业大学微生物防治省重点实验室,合肥230036 [2]安庆师范学院生命科学系,安庆246011

出　　处：《安徽农业大学学报》2010年第2期224-228,共5页Journal of Anhui Agricultural University

基　　金：国家"863"项目(2006AA10A212);安徽省优秀青年科技基金(08040106902);安徽省级自然科学研究重点项目(TD200708)共同资助

摘　　要：粉棒束孢是一种常见的昆虫病原真菌,被广泛用于生物防治。运用PCR技术与DNA步移技术,从粉棒束孢中克隆出类枯草杆菌蛋白酶的结构基因及其上游序列。结构基因总长1 650 bp,扩出的结构基因DNA与其cDNA比较在距离起始密码子369 bp处有一个内含子,大小为51 bp。通过DNA步移技术得到的上游序列大小为3 189 bp,与cDNA相互重叠的部分为352 bp。分析表明,上游序列中没有明显的TATA-盒和CAAT-盒,但含有GC-box、热激转录因子(HSF)等重要的转录因子结合位点,以及GATA等启动子顺式调控元件。Isaria farinosa is one of the most common species of entomopathogenic fungi,and has been used as biocontrol agents against some pests.The structural gene of subtilisin-like protease,with 1650 bp,was cloned from Isaria farinosa,and its upstream sequence was amplified by using genome walking.The amplified structural gene has a 51 bp intron compared with its cDNA.The obtained upstream sequence was 3189 bp,and overlapped 352 bp with cDNA.Analyses indicated that the upstream sequence contained several regulatory elements,such as GC-box,HSF and GATA,but the TATA-box and CAAT-box could not be found.

关 键 词：虫生真菌 粉棒束孢 基因步移 上游调控序列 

分 类 号：S476.12[农业科学—农业昆虫与害虫防治]