表达HSV-TK痘苗病毒真核表达载体pMJ601的构建与鉴定

Construction and Identification of Expression Vector pMJ6O1 Expressing HSV-TK Vaccinia Virus

目的：构建含自杀基因（ＨＳＶ－ＴＫ）的痘苗病毒真核表达载体ｐＭＪ６０１，为进一步实施胃癌的基因治疗作必要准 备。方法：利用目的基因与载体的连接、感受态细胞的制备及转化、质粒抽提、琼脂糖凝胶电泳、酶切等多种基因工 程技术将胶纯化回收的ＨＳＶ－ＴＫ与ｐＭＪ６０１进行连接、转化及鉴定。结果：克隆在Ｘ－ｐＰＮＴ质粒上的ＨＳＶ－ＴＫ ＤＮＡ 成功地被克隆到经ＢａｍＨⅠ－ＨｉｎｄⅢ双酶切的ｐＭＪ６０１载体上。结论：重组ＨＳＶ－ＴＫ痘苗病毒真核表达载体ｐＭＪ６０１ 的构建，为胃癌自杀基因的基因治疗打下坚实的基础。

Aims: To construct vaccinia expression vector pMJ601, containing HSV-TK for gene therapy of gastric cancer. Methods: Genetic engineering techniques as ligation, preparation of competent cell and transformation, plasmid extraction, agarose gel electrophoresis, restriction analysis were used for HSV-TK gene cloned to pMJ6O1 in the BamH I -Hind Ill multiple cloning site. Results: HSV-TK DNA fragment from the X-pPNT with restriction enzymes digest were shown on agarose gel electrophoresis. In addition, HSV-TK DNA was successfully cloned to pMJ6O1. Conclusions: It is one of the crucial steps for successful construction of pMJ6O1 expressing HSV-TK gene as a firm basis for further gene therapy of gastric carcinoma.