单纯疱疹病毒Ⅰ型胸苷激酶基因 Stocker 株的分子克隆及序列分析

Molecular Cloning of Thymidine Kinase Gene of Herpes Simplex Virus Type Ⅰ Strain Stocker and Its Sequence Analysis

根据已发表的单纯疱疹病毒 Ｉ型（ Ｈ Ｓ Ｖ Ｉ） Ｃ Ｌ１０１ 株胸苷激酶（ Ｔ Ｋ） 基因的核苷酸序列，设计并合成一对引物，以 Ｈ Ｓ Ｖ Ｉ Ｓｔｏｃｋｅｒ 株核酸为模板，应用 Ｐ Ｃ Ｒ 技术扩增出１ ．４２７ Ｋｂ 大小的片段，并将此片段克隆至载体ｐ Ｕ Ｃ１１９ 中，通过酶切和序列分析证明含完整的 Ｔ Ｋ 基因序列，此序列与 Ｃ Ｌ１０１ 株 Ｔ Ｋ 基因的核苷酸序列一致性为９９０ ％ ，氨基酸的一致性为９７６ ％ ，此基因的成功克隆为进一步研究 Ｈ Ｓ Ｖ Ｔ Ｋ Ｇ Ｃ Ｖ 系统在肿瘤基因治疗中的应用打下了基础。

A 1427bp DNA fragment containing thymidine kinase gene of herpes simplex virus type Ⅰ(HSV I) strain Stocker was amplified using primers based on TK gene sequence of HSV I strain CL101,and cloned into plasmid pUC119.Restriction enzyme digestion and sequence analysis indicated a complete TK gene in p119-TK,a 99.0% identity in nucleotides and a 97.6% identity in amino acids between the cloned TK gene and that from HSV I CL101.Cloning the HSVI TK laid the foundation for further study of tumor gene therapy with HSVI TK/GCV system.