单纯疱疹病毒胸苷激酶基因真核表达载体的构建及其在食管癌细胞中的表达  

Construction of eukaryotic expression vector containing thymidine kinase gene and its expression in esophageal cell line Eca-109

作　　者：李杰 郭玉忠[1] 谢华[1] 薛乐勋[1] 

机构地区：[1]郑州大学细胞生物学研究室,郑州450052

出　　处：《郑州大学学报（医学版）》2004年第1期47-50,共4页Journal of Zhengzhou University(Medical Sciences)

基　　金：河南省重大科技攻关基金资助项目 0 12 2 0 3 2 5 0 0 ;河南省杰出人才创新基金资助项目 0 2 2 10 0 190 0

摘　　要：目的 :构建含单纯疱疹病毒胸苷激酶 (HSV -tk)基因的真核表达载体 (pcDNA3-tk) ,并观察能否在食管癌细胞株中表达。方法 :将HSV -tk的cDNA亚克隆于真核表达载体pcDNA3上 ,构建pcDNA3-tk。后者经脂质体介导转染食管癌细胞株Eca - 1 0 9,通过高剂量的G4 1 8选择培养 ,筛选出抗性克隆。经RT -PCR检测确定HSV -tk基因是否能在转染细胞内转录。结果 :成功地构建了真核表达载体pcDNA3-tk ,并且在pcDNA3-tk转染的食管癌细胞内发现tk基因的mRNA。结论 :含HSVAim: To construct a recombinant eukaryotic expression vector containing HSV-tk cDNA and to investigate the expression of the vector in transfected esophageal cancer cell lines. Methods: pcDNA3-tk established by subcloning HSV-tk cDNA into a eukaryotic expression vector pcDNA3 was introduced into esophageal cancer cell lines Eca-109 by lipofectamine. The transfected esophageal cancer cells were selected in RPMI1640 containing G418, and RT-PCR was used for detection of mRNA transcription. Results: It was proved that the eukaryotic expression vector pcDNA3-tk was successfully constructed and HSV-tk mRNA was detected only in pcDNA3-tk transfected cell lines. Conclusion: pcDNA3-tk harboring HSV-tk cDNA can be expressed in the transfected esophageal cancer cells.

关 键 词：食管肿瘤 基因治疗 TK基因 单纯疱疹病毒胸苷激酶 真核表达载体 

分 类 号：R735.1[医药卫生—肿瘤]