摘要
目的克隆苏云金芽孢杆菌以色列亚种的杀蚊毒素蛋白基因cry11Aa,并使其在酵母双杂交系统受体菌Saccharomyces cerevisiaeL40中获得表达。方法应用聚合酶链反应扩增获得cry11Aa基因,构建表达载体pHybLex/Zeo-cry11Aa,转化到酵母菌Saccharomyces cerevisiaeL40。结果用Cry11Aa抗体和LexA抗体进行的Western blot免疫杂交表明cry11Aa基因在酵母菌L40中成功表达,生成了Cry11Aa-LexA融合蛋白。结论成功地克隆、表达了cry11Aa基因,为进一步利用酵母双杂交系统寻找与毒素蛋白Cry11Aa特异性结合的蚊幼中肠受体蛋白,揭示苏云金芽孢杆菌毒杀蚊虫的分子生物学机制奠定了基础。
Objective Clone of the mosquitocidal toxin gene cry11Aa from Bacillus thuringiensis subsp, israelensi and expression of this gene in the Yeast Two-Hybrid System Saccharomyces cerevisiae L40. Methods Gene cry11Aa was produced by PCR amplification. Expression vector pHybLex/Zeo-cryl 1Aa was constructed and transferred into Saccharomyces cerevisiae IAO. Results Western blotting with CryllAa and LexA antibodies showed that gene cryllAa was expressed in Saccharomyces cerevisiae IAO and a Cry11Aa-LexA fusion protein was produced. Conelusion Gene cry11Aa was successfully cloned and expressed. This work lay a foundation for searching the receptor in mce, quito larvae midgut which can interact with CryllAa and knowing more about the molecular biological mosquitocidal mechanism of Bacillus thuringiensis subsp, israelensis.
出处
《中国媒介生物学及控制杂志》
CAS
CSCD
北大核心
2006年第4期282-285,共4页
Chinese Journal of Vector Biology and Control
基金
国立墨西哥大学博士后基金
