【摘要】 目的克隆突变型单纯疱疹病毒胸苷激酶基因HSV1-sr39tk并构建其真核表达载体。方法通过PCR扩增HSV1-sr39tk基因,用BamHⅠ和BglⅡ双酶切纯化的PCR产物,在T4DNA连接酶作用下于室温将其连接于经BamHⅠ单酶切的慢病毒载体转移质粒pTK151上,重组质粒经PstⅠ酶切及测序鉴定。结果阳性重组质粒酶切及测序鉴定与预期结果完全相符。结论成功克隆出突变型自杀基因HSV1-sr39tk,并成功构建其真核表达载体。更多还原

【Abstract】 Objective To clone mutant herpes simplex virus thymidine kinase HSV1-sr39tk gene and construct its eukaryotic expression vector. Methods HSV1-sr39tk gene was amplified by PCR. PCR products were digested with BamHⅠand BglⅡ and inserted into lentiviral vector transfer plasmid pTK151 having been digested with BamHⅠin the presence of T4 DNA ligase at room temperature. The constructed recombinant plasmids were identified by digestion with PstⅠ and DNA sequencing. Results The outcome of restriction endonuclease digestion of positive recombinant plasmids and DNA sequencing were in consistence with the expected. Conclusion The mutant suicide gene HSV1-sr39tk gene was successfully cloned and eukaryotic expression vector carrying HSV1-sr39tk were constructed successfully.更多还原