构建单纯疱疹病毒Ⅱ型胸苷激酶基因真核表达载体

从单纯疱疹病毒Ⅱ型（HSV－Ⅱ）Sav株感染的Hep-2细胞培养液和细胞裂解液中提取HSV－Ⅱ基因组DNSA，以此为模板，用PCR方法获取胸苷激酶（TK）基因，将获取的DNA片段克隆入真核表达载体pcDNA3中，筛选阳性克隆测序，结果表明：本研究获取的HSV－Ⅱ－TK基因全部编码区为1128bp，编码376个氨基酸，结果表明：本研究扩增出HSV－ⅡTK基因的全部编码区序列，并成功构建真核表达载体pcDNA3／TK.

The Cloning of the Thymidine Kinase Gene of Herpes Simpler Virus II and the Constructing of Its Eukaryotic Expression Vector

The Hep-2 cells were infected by HSV-II Sav strain. HSV-II genomic DNA was purified from the Hep-2 cell culture medium and cell lysate and used as template to gain TK gene by PCR technique. The PCR product was cloned into eukarytic expression vector, pcDNA3 and sequenced. The sequencing date showed that coding region of HSV-II TK gene was 1?128 bp long coding for 376 amino acids. The TK gene cloned in this study was a full length sequence of coding region of HSV-II TK. The eukaryotic expressiong vector, pcDNA3/TK was constructed.