山羊ATF6基因重组慢病毒干扰载体的构建及鉴定

Construction and Authentication of Recombinant Lentiviral Vector for Goat ATF6 Gene Interference

为了研究ATF6通路介导的山羊胎盘滋养层细胞(goat trophoblast cell,GTC)凋亡的作用机制,需要构建激活转录因子6(ATF6)基因慢病毒干扰载体用于试验。采用RNA干扰技术,设计合成以山羊ATF6基因CDS区为靶点的shRNA,构建重组慢病毒载体、慢病毒包装、慢病毒转染、RT-qPCR和Western blot等方法,筛选出干扰效率达60%的shRNA干扰片段,构建出可用于今后ATF6通路相关研究的重组慢病病毒干扰载体,为山羊妊娠相关研究提供材料。

In order to research the mechanism of goat trophoblast cell (GTC) apoptosis by ATF6 gene mediated pathway,recombinant lentivirus vector of ATF6 gene interference needs to be constructed.In this experiment,we designed and synthesised the ATF6-targeted shRNA,constructed the recombinant plasmid,packaged the ATF6-interference lentivirus and transfected it in goat endometrial epithelium cell (EEC),then used qRT-PCR and Western blot to filtered out the ATF6-targeted shRNA which interference rate up to 60%,constructed the recombinant lentivirus vector for goat ATF6 gene interference,that can be helpful for studies on goat pregnancy in the future.