PCR点突变改造人透明带蛋白3酵母表达载体

Modification of yeast expression vector of human pellucid zone protein 3 by point mutation of PCR

目的:构建人透明带蛋白3的186位氨基酸突变体重组质粒并鉴定。方法:通过PCR点突变改造人透明带蛋白3的Kex2位点,构建pPICZα-hZP3186S突变载体。结果:经DNA测序表明,ZP3基因的556～558bp处碱基由AGG突变为TCC,读码框正确。结论:成功构建了pPICZα-hZP3186S突变载体(186位精氨酸突变为丝氨酸),为重组人ZP3蛋白在毕赤酵母能够分泌性表达奠定基础。

Objective：To construct and identify a recombinant plasmid of human pellucid zone protein3 （ZP3） containing186 amino acid mutants.Methods：Kex2 site of ZP3 was modified by point mutation of PCR,pPICZα-hZP3 186S mutant vector was constructed.Results：DNA sequencing showed that AGG of 556 ～ 558bp sites in ZP3 gene changed into TCC,and the reading frame was correct.Conclusion：pPICZα-hZP3 186S mutant vector is constructed successfully,which provides a foundation for the expression of human ZP3 protein in Pichia pastoris.