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Position: Home > Articles > Cloning and expression of the receptor gene (Bt-r_3) of the Bt toxic protein Cry1Ab in E. coli. Journal of Zhejiang University (Agriculture and Life Sciences) 2004,30 (3)

Bt毒蛋白Cry1Ab受体基因Bt-r_3在大肠杆菌中的克隆和表达及特性分析

作  者:
徐步进;徐强;吴志平;华跃进;陈子元
单  位:
浙江大学原子核农业科学研究所
关键词:
Cry1Ab;受体基因;Bt-r3;克隆;表达
摘  要:
根据二化螟中肠Bt毒蛋白Cry1Ab的受体基因Bt-r3的cDNA序列设计特异引物,扩增得到编码Bt-R3蛋白胞外部分的目的片段(约4500bp).将目的基因酶切克隆到质粒载体prcHis-TOPO上,再将该质粒导入到感受态细胞TOP10,经IPTG诱导、得到约为160kD的表达蛋白.经Western印迹鉴定,结果表明表达产物能有效地结合Bt毒蛋白Cry1Ab,从而确认Bt-r3是Cry1Ab的受体基因.它们的结合部位位于Bt-R3蛋白的胞外结构域中.
译  名:
Cloning and expression of the receptor gene (Bt-r_3) of the Bt toxic protein Cry1Ab in E. coli.
作  者:
XU Bu-jin, XU Qiang, WU Zhi-ping, HUA Yue-jin, Chen Zi-yuan (Institute of Nuclear-Agricultural Sciences, Zhejiang University, Hangzhou 310029, China)
关键词:
Cry1Ab; receptor gene; Bt-r_3; clone; expression
摘  要:
An extracellular part of the receptor gene (Bt-r_3) of the Bt toxic protein Cry1Ab, with a length of about 4500 bp, was amplified by PCR and cloned into plasmid prcHis-TOPO. The recombinant plasmid pTrcHis-TOPO/peBt-r_3 was transformed to E. coli, and expressed at 37 ℃ by IPTG induction. SDS-PAGE proved that the molecular weight of expressed product was about 160 kD. Western blotting showed that the recombinant protein could specifically bind to Bt toxic protein Cry1Ab, which validated that the Bt-R_3 was a receptor protein of Cry1Ab. The binding site of Bt-R_3 with Cry1Ab was located in extracellular domains of Bt-R_3 protein.

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