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

作　　者：王弘珺[1] 李质馨[1] 徐冶[1] 田洪艳[1] 朱辛为[1] 潘晓燕[1] 窦肇华[1] 

机构地区：[1]吉林医药学院组织胚胎学教研室,吉林吉林132013

出　　处：《中国妇幼保健》2010年第29期4282-4283,共2页Maternal and Child Health Care of China

基　　金：吉林省科技发展计划资助项目(200705420)

摘　　要：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.

关 键 词：透明带 点突变 PCR 毕赤酵母 

分 类 号：R321.1[医药卫生—人体解剖和组织胚胎学]