绵羊FecB基因打靶载体构建

Construction of a new gene targeting vector in the sheep

利用peGFP-C1,pRET.IL.PGK-TK和pMD19-T Simple等基本质粒,构建绵羊FecB基因打靶载体。以克隆载体pMD19-T Simple为载体骨架,插入一个正选择标记eGFP基因、一个负选择标记基因HSV-tk基因,并在eGFP基因两侧各添加一个同源长臂和同源短臂,从而构建绵羊FecB基因打靶载体。结果:经多个限制性内切核酸酶酶切鉴定和测序证实,所构建的基因打靶载体符合设计要求,表明成功构建了绵羊FecB基因打靶载体。

In order to construct a sheep FecB gene targeting vector, plasmids of peGFP-C1, pRET. IL. PGK-TK and pMD19-T Simple and pMD19-T Simple cloning vector as the carrier frame with a positive selection marker eGFP gene and a negative selection marker gene HSV-tk gene were used. The correct structure of the targeting vector was confirmed by restriction enzyme digestion and sequencing analysis. Our experiment indicated that a FeeB gene targeting vector of sheep was successfully constructed.