用昆虫病毒转移载体在大肠杆菌中表达苏芸金杆菌δ-内毒素基因  

Expression of Endotoxin Gene from Bacillus thuringiensis with Insect Baculovirus Transfer Vector in Escherichia coli

作　　者：裴子飞[1] 齐义鹏[1] 黄永秀[1] 沈英[2] 

机构地区：[1]武汉大学病毒学系 [2]同济医科大学微生物教研室

出　　处：《生物工程学报》1993年第2期181-183,共3页Chinese Journal of Biotechnology

基　　金：国家自然科学基金;国家教委博士点基金

摘　　要：从八十年代初开始用昆虫核型多角体病毒(NPV)的多角体蛋白基因(ocu)构建转移载体以来,陆续表达了几十种外源基因。我们曾用大尺蠖核多角体病毒(BsNPV)的ocu基因构建了转移载体。After a 6.6kb fragment carried δ-endotoxin gene of Bacillus thuringiensis kurstaki HD-73 was obtained from plasmid pBt28 by HindⅢ, it was subcloned into downstream of polyhedrin gone promoter in baculovirus transfer vector pBsA39. The δ-endotoxin gene as inserting fragment is present in chimeric vector by Southern blot hybridizition.The δ-endotoxin gone on the chimeric vector pBsBt70 etc. is able to express two proteins with MW of 68 and 23 kDa and yield of 16.257μg/ml and 11.335μg/ml respectively in E.coli cells. Expression proteins from chimeric vector are able to immunoprecipitate with antibody prepared by purified HD-73 crystal proteins. Upon biological toxicity test to cabbage catorpilar by lysate, the expression proteins were toxin with a mortality of 100% on 6th day to the 3 instar larvae. It is demostrated that the expression proteins wbich probably are decomposational products from pretoxin with MW of 138kDa, or 90kDa, are Bt HD-73 crystal protein with biological function.

关 键 词：苏芸金杆菌 生物杀虫剂 内毒素基因 

分 类 号：S188[农业科学—农业基础科学]