用点突变PCR方法去除人IL－2cDNA表达克隆终止密码及其鉴定

Deleting stop Codon of Human IL-2 cDNA Expression Clone through Side Mutation PCR and Identification

在重组人ＩＬ－２（ｈＩＬ－２）大肠杆菌高效表达的基础上，为了构建ｈＩＬ－２与别的目的蛋白的嵌合分子，我们利用点突变ＰＣＲ方法去除ＩＬ－２ｃＤＮＡ表达克隆的终止密码，并构建成ｈＩＬ－２嵌合重组蛋白表达克隆，在大肠杆菌中表达的ＩＬ－２嵌合重组蛋白具有天然ＩＬ－２分子的生物活性。本文结果表明点突变ＰＣＲ去除ｃＤＮＡ终止密码的方法效率高、快速、简便。

On the basis of over-expression of recombinant hIL-2 in E. coli,in order to construct multifunction chimeric molecules of IL-2 and other interest proteins,we deleted the stop codon of hIL-2cDNA expression clone through site mutation directed by PCR. The hIL-2 cDNA without stop codon was recombined with expression vector pKPL,constructed into expression clone pKPL-mhIL-2, which could over-expressed a chimeric recombinant protein of IL-2.The results here showed that the side mutation PCR method was rapid, convenient and high efficient for deleting stop codon of cDNA.