峨边花牛肌肉生长抑制素基因cDNA克隆及序列分析

目的]为改良峨边花牛地方品种、提高产肉率及种质资源的保存奠定基础和提供技术依据。方法]以从乐山峨边花牛骨骼肌中提取总RNA为模板,利用RT-PCR技术扩增获得肌肉生长抑制素基因(MSTN)cDNA片段,将其克隆到T载体上,通过PCR鉴定阳性克隆并对其进行测序鉴定。结果]通过PCR扩增,得到峨边花牛MSTN基因cDNA的全序列,全长1 135 bp,包含全部的MSTN编码序列,共有3个外显子。将峨边花牛MSTN基因在GenBank上与比利时双肌牛及皮埃蒙特双肌牛比较后发现,峨边花牛在上述2种品种双肌牛的突变位点碱基未发现异常。结论]成功地克隆峨边花牛的MSTN基因并对其进行测序对于后期表达载体的构建奠定了基础。

cDNA Cloning and Sequence Analysts of Myostatin Gene from Ebian Flower Cattle

Objective] The aim was to lay the foundation and provide the technical basis for improving the local varieties of Ebian flower cattle,advancing the meat production rate and preserving germplasm.Method] The cDNA fragment of myostatin(MSTN) gene was amplified from the total RNA extracted from the Ebian flower cattle muscle by RT-PCR technique,then inserted into T vector and then sequenced after screening the positive clones identified with PCR.Result] A cDNA full sequence of MSTN gene from Ebian flower cattle muscle was amplified by PCR and its full length including the complete coding sequence of MSTN gene was 1 135 bp,which had 3 exons.Compared with the MSTN gene from Belgium double muscle bovine and Piedmont double muscle bovine in GenBank,the MSTN gene from Ebian flower cattle had no anomaly in the bases of mutation site of the MSTN gene from the above double muscle bovine varieties.Conclusion]The MSTN gene from Ebian flower cattle was cloned successfully and sequenced,which laid the foundation for the construction of expressive vector in the latter research.