摘要
本研究根据Genbank中牛肌肉生成抑制素基因序列设计引物,在设计的引物两端分别加上限制性内切酶BamHI和EcoRI的识别位点序列。利用RT-PCR技术从西门塔尔牛肌肉组织的总RNA中扩增出MSTN基因的CDNA序列,扩增出1 125 bp片段,该片段与pMD18-T载体连接,转化JM109感受态细胞,所得阳性克隆进行酶切和PCR鉴定,并进行了测序分析,得到的克隆序列与设计的序列基本一致,表明成功地克隆了西门塔尔牛的肌肉生成抑制素基因的蛋白编码序列。
The primer added BamH Ⅰ and EcoR Ⅰ recognition site sequence were designed according to the cattle myostatin gene sequence in the Genbank. 1 125 bp fragment of myostatin gene maturation region was amplified by RT-PCR from muscular tissue of Simmental cattle, and ligated with vector pMD18-T and transfeeting JM109 competent cell. The sequence of identified positive done by PCR and digested with restriction enzyme was analyzed, the same sequence between clone and designed sequence indicated that the myostatin gene of Simmental cattle was cloned successfully.
出处
《中国牛业科学》
2006年第1期19-20,23,共3页
China Cattle Science
基金
国家自然科学基金资助项目(项目编号:30500366)
