【摘要】 目的　构建人血栓调节蛋白 (humanthrombomodulin ,hTM )基因的真核表达质粒 ,为研究hTM在抗血栓、抗炎症、抗血管增生等方面的作用及其临床应用奠定基础。方法　用PCR法扩增出hTMcDNA表达片段。把载体质粒 pcDNA3.1 (+) /neo和hTM片段用EcoRI +HindⅢ双酶切 ,T4DNA连接酶进行连接。结果　重组质粒在大肠杆菌菌株DH5α内扩增。提取、纯化后用HindⅢ、EcoRI酶切鉴定及测序鉴定证明hTMcDNA表达片段已被完整、正确的插入到pcDNA3.1 (+) /neo质粒中。 结论　成功构建了hTM基因的真核表达质粒pcDNA hTM。更多还原

【Abstract】 Objective To construct eukaryotic expression plasmoid of human thrombomodulin gene for further study on hTM function on anticoagulant ,antiflammatoyr and neointima formation resistant.Methods The hTM gene fragment was amplified by polymerase chain reaction(PCR)from human great saphenous vein.Then,both pc DNA3.1(+)neo vector and the hTM fragment were digested by hindⅢ/EcoR I.Two digested fragments were ligated with T 4DNA ligase and pcDNA hTM plasmid was formed.Results After the identification of redigesting and sequencing,the reconstructive plasmid was confirmed which contained the correct and full nucleotide sequence of hTMcDNA in pcDNA3(+)/neo.Conclusion The pcDNA hTM plasmid was consructed.更多还原