小菜蛾化学感受蛋白突变体的构建

摘要

[Objective] This study aimed to explore the effects of cysteine (Cys58) on the characteristics of PlxyCSPl in P. Xyostella combined with pesticide compounds. [Method] Cys58 of PlxyCSPl was mutated into Trp58 by using overlap extension PCR method, and PlxyCSPl -M2 mutant was obtained. Expression vector was constructed and the protein was detected by western blot. [Result] Expression vector pET32a- PlxyCSPl -M2 was constructed to express the 35 kDa weight protein. [Conclusion] PlxyCSPl mutant protein was expressed successfully in prokaryotic expression system.%[目的]研究半胱氨酸(Cys58)对小菜蛾(Plutella xylostella)化学感受蛋白1(PlxyCSP1)与农药化合物结合特性的影响.[方法]利用重叠延伸PCR法,将PlxyCSP1中Cys58突变成色氨酸(Trp58),获得突变体PlxyCSP1-M2.构建表达载体,表达突变蛋白,并进行Western Blot检测.[结果]构建了突变基因的原核蛋白表达载体pET32a-PlxyCSP1 -M2,并表达了大小为35 kDa的融合蛋白.[结论]利用重叠延伸PCR法成功在原核系统中表达了PlxyCSP1突变蛋白.