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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DNA ,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 :本研究所获取的HSV -Ⅱ -TK基因全部编码区为 112 8bp ,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 .
HSV-II genomic DNA was extracted from Hep-2 cell culture and cell lysate infected with HSV-II strain Sav strain, and was used as a template to obtain thymidine kinase (TK) gene by PCR. The obtained DNA fragment was cloned into the eukaryotic expression vector pcDNA3 and the positive clones were screened.The results showed that the entire coding region of HSV-Ⅱ-TK gene was 112 8bp encoding 376 amino acids.The results showed that: The complete coding sequence of HSV-ⅡTK gene was amplified and the eukaryotic expression vector pcDNA3 / TK was successfully constructed.