用于分子改造的苏云金芽胞杆菌cry1Ab基因的克隆与表达  

Cloning and Expression of cry1AbGene from Bacillus thuringiensis for Further Molecular Modification

作　　者：陈凡冰[1,2] 邵恩斯[1,3] 黄志鹏[1] 

机构地区：[1]福建农林大学教育部生物农药与化学生物学重点实验室,福州350002 [2]福建农林大学生命科学学院,福州350002 [3]福建农林大学植物保护学院,福州350002

出　　处：《生物技术进展》2013年第2期132-136,157,共5页Current Biotechnology

基　　金：国家自然科学基金项目(31071745);福建省自然科学基金项目(2011J01076)资助

摘　　要：苏云金芽胞杆菌(Bacillus thuringiensis)cry1Ab基因已广泛用于抗虫转基因植物,但其编码的杀虫蛋白对刺吸式口器害虫基本无效。本研究依据已发表cry1Ab基因序列设计一对全长引物,从BtWB7菌株总DNA中克隆出cry1Ab基因全序列,构建原核表达载体pGEX-KG-cry1Ab并转化到大肠杆菌BL21(DE3)中,经IPTG诱导成功表达出156kDa的Cry1Ab-GST融合蛋白。研究结果为进一步定向改造Cry1Ab使其能够正确识别刺吸式口器害虫肠道内的受体蛋白奠定基础。The crylAb gene from Bacillus thuringienzis has been widely used in transgenic plants around the world. However, the Cryl Ab insecticidal protein shows no or little toxicity to the pests with piercing-suckling moutnparts. In this study, the crylAb gene was amplified from total DNA of Bt strains WB7 with a pair of primers designed on the full-length sequences of published crylAb genes. Then it was ligated with linearized pGEX-KG vector to construct recombinant expression vector pGEX-KG-crylAb. The soluble CrylAb-GST fusion protein was obtained after transferring pGEX-KG-crylAb into E. coli BL21 （ DE3 ） and then inducing by IPTG. The results provided a basis for further studies of directional modification of CrylAb in order that it can bind to the proper receptors in midgut of the pests with piercing-suckling moutnparts.

关 键 词：苏云金芽胞杆菌 CRY1AB基因 克隆与表达 可溶性蛋白 

分 类 号：S476.1[农业科学—农业昆虫与害虫防治]