羊卵泡抑制素α亚基基因siRNA真核表达载体的构建  

Construction of siRNA expression vector targeting inhibin α subunit gene of sheep

作　　者：黄增文 木尔扎提 郑婷婷[1] 罗玉江[1] 赛务加甫[1] 吾热力哈孜.哈孜汗 

机构地区：[1]石河子大学动物科技学院,新疆石河子832003

出　　处：《畜牧与兽医》2016年第4期9-14,共6页Animal Husbandry & Veterinary Medicine

基　　金：石河子大学科学技术研究发展计划项目(gxjs2011-yz09)

摘　　要：为了构建羊抑制素α亚基(IHNα)基因靶向siRNA真核表达载体,以也木勒白羊为试验动物,依据Gen Bank的INHα基因序列(KP-113678.1)设计3个不同位点的干扰片段,利用RNAi技术和实验室常规方法将3个干扰片段连接到干扰RNA真核表达载体中,并对干扰载体进行相关检测和鉴定。结果表明:干扰表达质粒能在绵羊颗粒细胞中成功表达,且转染率50%左右,通过Q-PCR和蛋白表达试验发现,沉默率达到83%。In order to construct siRNA expression vector targeting inhibin α subunit（ IHNα） gene,three different RNAi sites were designed according to the INH α gene sequence（ Gen Bank： KP-113678. 1）. We used RNAi technology and routine laboratory methods to ligate siRNA into the eukaryotic expression vector of RNA interference,and RNAi vector were confirmed by the related detection. The results showed that the plasmid was successfully expressed in sheep granulosa cells,and the transfection rate was around 50%. The siRNA silence rate reached 83% as showed by Q-PCR and protein analysis experiment. Our results will lay the foundation for developing transgenic cell model of Yemil Aries INH gene interference.

关 键 词：也木勒白羊 INHα RNA干扰 PCR技术 荧光定量PCR 

分 类 号：S826[农业科学—畜牧学]