叶螨AChE突变型的原核表达及活性测定

Prokaryotic expression and enzyme activity analysis of acetylcholinesterase mutants from Tetranychus cinnabarinus

【目的】利用朱砂叶螨AChE突变型来寻找防治农业主要害螨的选择性杀螨剂靶点。【方法】构建螨AChE突变型原核表达载体,纯化蛋白后进行酶活性比对分析。【结果】AChE突变型于pET-30a载体中成功大量表达,获得2个较高纯度的AChE突变型重组蛋白AChE-VWE和AChE-WEH,它们与野生型相比虽活性降低,但差异不显著。【结论】实现了对AChE突变型原核高效表达系统构建,获得AChE突变型蛋白,为螨AChE功能研究及新型选择性杀螨剂的筛选奠定基础。

【Objective】The acetylcholinesterase mutants of Tetranychu scinnabarinus were used to seek the target of selective acaricide to control the main pest mite in agriculture.【Methods】The expression vectors of AChE mutants of Tetranychus cinnabarinus were constructed,and the proteins were purified.Moreover,their enzyme activities were comparatively analyzed.【Results】The AChE mutants were successfully expressed in pET-30a vector,and high quality of the recombinant proteins were purified.Compared with the wild type AChE,the activities of the AChE-VWE and AChE-WEH variants were lower,but the difference was not significant.【Conclusion】The high efficient prokaryotic expression system of AChE mutants were constructed,and AChE mutant proteins were obtained,which provides the basis for the exploration of AChE function and structure,and for the screening of new selective acaricide.