PIWIL1基因启动子调控的特异表达Cre重组酶的转基因猪的构建

Construction of Transgenic Swine Specifically Expressing Cre Recombinase Controlled by PIWIL1 Promoter

[目的]建立PIWIL1基因启动子调控的特异性表达Cre重组酶的转基因猪,为研究piRNA及其相关基因的功能和机制奠定基础。[方法]通过PCR扩增,从猪的全基因组上扩增出PIWIL1基因的启动子。切除猪源Cre重组酶的表达载体pET28a-MHC-Cre-BGHpo1yA-FRT2neor的启动子MHC,将其与PIWIL1基因启动子连接,构建成特异性表达Cre重组酶的pPIWIL1-Cre载体。将该载体转染小型猪胎儿成纤维细胞,通过G418筛选,获得阳性克隆进行核移植。[结果]获得2头PIWIL1基因启动子调控的特异表达Cre酶的转基因猪。[结论]成功构建了PIWIL1基因启动子调控的特异表达Cre重组酶的转基因猪,为研究piRNA及其相关基因的功能和机制提供了工具猪。

[Objective] The aim was to generate transgenic pig specifically expressing Cre recombinase controlled by PIW1L1 promoter and make basic contribution for research on piRNA and its related genes＇ function and mechanism. [ Method] The promoter of pig PIWIL1 gene was amplified by PCR method. Replace the MHC promoter of pig specific Cre recombinase expression plasmid （pET28a-MHC-Cre-BGH- polyA-FRT2neor） with PIWIL1 promoter and specific Cre recombinase plasmid （pPIWIL1-Cre） was obtained. Transfect the plasmid to minipig fetal fibroblast cells and screen for positive clones with drug and do nuclear transfer. [ Result ] Two transgenic pigs of specific expressing Cre recombinase controlled by PIWILI promoter were got. [ Conclusion] We generate transgenic pigs that specifically express Cre recombinase successfully and make ~ossible tools for piRNA and related gene function and mechanism research.