含TKglyES融合基因重组质粒的构建

目的：构建携带胸腺嘧啶激酶（thymidine kinase，TK）及内皮抑素（Endostatin，ES）融合基因的重组质粒。方法：以pAdE1CMV—TK为模板，经聚合酶链反应（PCR）扩增TK片段，将其插入腺病毒穿梭质粒pAdTrack—CMV，再以PET-19b—ES质粒为模板经PCR扩增glyES片段，将其插入TK片段的下游，构建成pAdTrackCMV—TKglyES。结果：酶切、PCR及DNA测序鉴定表明成功构建了携带胸腺嘧啶激酶（thymidine kinase）及内皮抑素（Endostatin）融合基因的重组质粒。结论：为进一步构建病毒载体用于肿瘤基因治疗打下基础。

Construction of recominant plasmid containing TKglyES fusion gene

Objective: To construct the recombinant plasmid containing thymidine kinase glycine endostatin fusion gene. Methods: The DNA encoding thymidine kinase was amplified by polymerase chain reaction(PCR) from the plasmid of pAdE1CMV-TK, then it was inserted into the shuttle plasmid of pAdTrack-CMV; the DNA encoing glycine and enostatin was amplified by PCR from the plasmid of PET-19b-ES. The fragment of glycine endostatin was inserted at the backward position of the TK fragment to construct the pAdTrackCMV-TKglyES. Results: Digestion, PCR and DNA sequencing indicated that the recombinant plasmid containing thymidine kinase glycine enostatin fusion gene was constructed successfully. Conclusion: It provide a basis for construction of virus recombinant which could be applied in the field of tumor therapy.