猪肌生成抑制素(MSTN)cDNA的克隆

Cloning of Swine Myostatin Gene cDNA

从猪的肌生成抑制素 ( MSTN)编码序列中设计引物 ,以军牧一号猪肌细胞总 RNA为模板 ,利用 RT-PCR和嵌套 PCR技术 ,扩增出 MSTN c DNA片段。该片段全长 1 2 77bp,包含猪 MSTN基因的全部编码序列。将所得片段与 p MD1 8-T载体连接 ,转化到 JM1 0 9大肠杆菌中 ,成功地筛选到阳性克隆 ,其质粒测序结果与文献报道的一致。经 Eco R 和 Pst 酶解分析 ,c DNA片段与 p MD1 8-T载体之间既有正向插入的克隆 ,也有反向插入的克隆 ,所得到的 MSTN c

Total RNA of muscle cell was prepared from a new pig breed JunMu 1 and used in RT PCR to amplify myostatin(MSTN) gene cDNA.The resulted cDNA fragment with the expected size was cloned into vector pMD 18 T and subsequently subjected to restriction enzyme analysis and sequencing. The result showed that the cloned MSTN gene is 1 277 bp comprising the complete coding sequence and 99.5% homogenous to published data with replacement of 4 nucleotides causing only one amino acid changed.The expression of cloned MSTN gene in prokaryotic and eukaryotic system is under way.