湖羊FecB基因在新疆细毛羊胎儿成纤维细胞中的表达

Expression of Hu Sheep FecB Gene in Fetal Fibroblasts of Xinjiang Fine-wool Sheep

为培育携带FecB多胎基因的高繁殖率新疆细毛羊,本研究提取了湖羊卵巢的总RNA,在此基础上反转录为cDNA,通过PCR扩增得到了湖羊的FecB基因,再与pMD19-T载体连接,构建了pMD19-T-FecB重组质粒。通过双酶切与真核表达载体pEGFP-N1连接,得到了pEGFP-N1-FecB真核表达载体。经测序与酶切证明,成功构建了pEGFP-N1-FecB真核表达载体,并转染新疆细毛羊胎儿成纤维细胞,通过药物筛选获得了表达湖羊FecB基因的阳性新疆细毛羊胎儿成纤维细胞,为培育高繁殖率的新疆细毛羊奠定基础。

To cultivate the Xinjiang fine-wool sheep with high reproduction rate, the ovarian total RNA of Hu sheep was extracted and reverse-transcribed into cDNA, and Hu sheep FecB gene was obtained by PCR amplification. Hu sheep FecB gene was linked to pMD19-T plasmid to construct the pMD19-T-FecB recombinant plasmid, pEGFP-N1-FecB eukaryotic expression vecor was constructed, and Hu sheep FecB gene was linked to pEGFP-N1 vector by double digestion. The sequencing and restriction enzyme digestion results showed that the pEGFP-N1-FecB eukaryotic expression vector was successfully constructed. The pEGFP-N1-FecB was thansfected into fetal fibroblast cells of Xinjiang fine-wool sheep and the positive cells was gotten by drug resistance. The cells gotten in this experiment can be used as the foundation to cultivate the Xinjiang fine-wool sheep with high reproduction rate.