摘要
从猪肌生成抑制素 ( MSTN) c DNA序列中设计引物 ,以 p MD18- T- MSTN质粒为模板 ,PCR扩增猪 MSTN成熟蛋白编码序列 ,该片段全长 10 95 bp。将所得片段与 p MD18- T载体连接 ,构建重组质粒 p MT- mp MSTN,将其转化到大肠杆菌 DH5α中 ,成功地筛选到阳性克隆。双酶切重组质粒 p MT- mp MSTN和真核表达载体 pc DNA3.1,连接目的片段与载体 pc DNA3.1,转化大肠杆菌 DH5 α,经酶切、PCR及测序分析 ,表明成功地构建了真核表达质粒 pc DNA3.1-mp MSTN。
In this research,the coding sequence of MSTN mature protein was amplified by PCR from myostatin plasmid pMD18 T MSTN of a new porcine breed JunMu 1.The fragment with the expected size was cloned into vector pMD18 T and subsequently subjected to PCR,restriction endonucleases analysis and sequencing.Then,the fragment was cloned into vector pcDNA3 1 for eukaryotic expression.The coding sequcene of MSTN mature protein was coincidence with our design after PCR,restriction endonucleases analysis and sequencing.The expression of the cloned coding sequence of MSTN mature protein in eukaryotic system is under way.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2003年第6期610-612,共3页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目 (3 0 0 70 5 63 )
军队医药卫生青年基金资助项目(98Q0 79)
