牛抑制素α亚基基因克隆及表达载体的构建

Cloning and Construction of Expressive Vectors of Bovine Inhibin α Subunit Gene

为克隆牛抑制素α亚基基因,构建抑制素p ET32a-成熟肽表达载体,从河北大厂县屠宰场采集卵巢,提取总RNA并反转录成c DNA,根据Gen Bank NM_174094.4设计一对引物扩增出成熟肽c DNA,将所扩增基因插入到p ET32a表达载体Bam HⅠ与HindⅢ酶切位点之间,构建重组质粒抑制素p ET32a-成熟肽。结果成功构建出抑制素α亚基基因。笔者初步得到了抑制素重组质粒,为今后重组蛋白原核表达的研究奠定了基础。

The research aimed to clone the bovine inhibin α subunit gene and construct inhibin p ET32a-matpeptide expression vector. The bovine ovaries were collected from slaughterhouses of Dachang County in HebeiProvince. The total RNA was extracted and reverse transcribed into c DNA. A pair of primers was designed byPrimer 5.0 according to the known bovine inhibin α-subunit sequence（Genbank No.NM174094.4）. Inhibin αsubunit mat peptide gene was amplified by RT- PCR. Finally, c DNA of this matured peptide was furtheramplified and cloned into the Bam H Ⅰ and Hind Ⅲ sites of p ET32 a expressing vector to generate therecombinant inhibin p ET32a-mat peptide plasmid. The results showed that the inhibin α subunit gene wassuccessfully constructed. The authors preliminarily obtained the recombinant plasmid of inhibin and laidfoundation for the further study of prokaryotic expression of recombinant proteins.