猪促卵泡激素α和β亚基基因的克隆及其重组腺病毒表达载体的构建

Cloning of α and β subunit genes encoding swine FSH and construction of recombinant adenovirus with α or β subunit gene

为探索人工制备猪促卵泡激素的可行性,从猪的脑垂体中抽提总RNA,通过RT-PCR方法,克隆了猪促卵泡激素α、β亚基基因,并将其分别定向插入腺病毒穿梭质粒中,通过转化BJ5183-AD-1细胞继而构建含有猪促卵泡激素α、β亚基基因的重组腺病毒质粒;将含有猪促卵泡激素α、β亚基基因的重组腺病毒质粒转染AD-293细胞,细胞产生病变,证实猪促卵泡激素α、β亚基基因重组腺病毒表达载体构建成功。

In order to explore the feasibility of artificial preparation of swine FSH,the total RNA was extracted from tissues of swine pituitary gland,and the α and β subunit genes encoding swine FSH were cloned by RT-PCR,and then inserted into the shuttle-CMV plasmid separately to construct recombinant adenovirus plasmids with α or β gene by transforming BJ5183-AD-1 cells.Then,homologous recombination with the α or β gene led AD-293 cells to be CPE,indicating that the recombinant adenovirus with α or β subunit gene encoding swine FSH was constructed successfully in vitro.