利用定点突变技术构建血型糖蛋白Ac末端突变体

Constructing mutant GPA c-terminus plasmids by the transformer site-directed mutagenesis technique

目的 构建血型糖蛋白A(GlycophorinA)c末端突变体 ,为GPAc末端与阴离子交换蛋白 1(AnionExchang er1,AE1)酸性c端域相互作用位点研究奠定基础。方法 以pM GPA Ct为基础质粒 ,利用TransformerTM定点突变技术 ,构建GPAc末端突变体pM GPA L118F、pM GPA S119R。结果 经过测序证实突变成功。结论 本实验成功构建了GPAc末端的突变体 ,并证实TransformerTM定点突变方法具有简便、突变效率高等优点 ,是体外构建突变体的有效方法。

Objective The mutant Glycophorin A c-terminus gene plasmids were constructed in order to study the function sites of GPA c-terminus.Methods Using Transformer TM site-directed mutagenesis,the mutant site was induced into the plasmid of pM-GPA-Ct.Results The mutant plasmids proved by sequencing were successfully constructed.Conclusion The mutant plasmids are used for the analysis of the function sites of GPA c-terminus.The transformerTM site-directed mutagenesis technique has the advantages of simplicity and high-efficiency.