猪IκBα基因定点突变中两种方法的应用  

Application of Two Methods for Site-specific Mutagenesis of Porcine IκBα

作　　者：李和刚[1,2] 傅田[1] 靳二辉[1] 操继跃[2] 李奎[1] 肖邦[1] 杨述林[1] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所中国农业科学院家养动物遗传资源与种质创新重点开放实验室,北京100193 [2]华中农业大学动物医学院,武汉430070

出　　处：《中国畜牧兽医》2009年第1期38-40,共3页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(30771547);“973”项目(2006CB102105);“863”项目(2006AA10Z135)

摘　　要：作者将猪IκBα基因32、36位丝氨酸位点突变为丙氨酸,为进一步研究突变型IκBα蛋白对异种移植延缓性排斥反应的抑制机理奠定基础。首先采用Stratagene公司单点突变试剂盒将猪IκBα基因32位丝氨酸位点突变为丙氨酸,接着试图在36位引入突变但未成功,最后改用重叠延伸PCR对36位进行突变。结果表明,两种突变方法结合使用,成功地将猪IκBα基因32、36位丝氨酸位点突变为丙氨酸。说明两种突变方法各有所长,又有各自的缺点,应根据试验的实际需要灵活选择合适的突变方法,以收到事半功倍的效果。Substitute Ala for the sites of Ser 32 and Ser 36 in porcine IκBα gene,to establish the fundament for mechanism research of the mutant IκBα inhibition on delayed xenograft rejection. QuikChange^R Site-Directed Mutagenesis Kit was first uesd for the mutagenesis of Set 32 site,then the kit was subsequently used for the mutagenesis of Ser 36 site,but failed. Finally OE-PCR was used for the mutagenesis of Ser 36 site. These two mutagenesis methods were combined to achieve site-specific mutagenesis at the sites of Ser 32 and Ser 36 in porcine IκBα gene. These two methods each has a unique, and each has their own shortcoming. So, in order to get better result by less work,mutation methods should be chosen flexibly according to the real need.

关 键 词：定点突变 重叠延伸PCR 猪IκBα基因 

分 类 号：Q754[生物学—分子生物学]