中文摘要：

背景自杀基因治疗是为头和颈癌症的广泛地使用的分子的治疗。在这研究，我们试着在细菌使用同质的再结合的方法克隆 thymidine kinase 基因( tk )到为复制有缺陷者 adenoviruses.Methods pAdTrack-CMV/tk 的构造的侵入人体气管粘膜的病菌脊梁向量的一种自杀基因从 plasmid pCMV-tk 包括 thymidine kinase 基因通过限制 endonuclease 碎片的 subclone 被构造到另一 plasmid pAdTrack-CMV ，然后有 supercoiled pAdEasy-1 的 co-transfected ，除了，它是 adenoviral 脊梁向量与一样的方法， pAdTrack-CMV 也在 BJ5183 为同质的再结合与 pAdEasy-1 被共同转变。与限制 endonuclease Pacl 和聚合酶链鉴别反应(PCR ) ， plasmids pAd-GFP/tk 和 pAd-GFP 成功地被构造。他们中的每与 Pacl 并且 sequently 被消化 transfected 进用 Lipofectamine 2000 .Results 的人的胚胎肾 293 房间(HEK293 ) Ad-GFP/tk 和 Ad-GFP 的像彗星的生产侵入人体气管粘膜的病菌的 foci 在 5 ～ 7 天房间 culture.After 以后被观察 12 天包装，复制有缺陷者侵入人体气管粘膜的病菌被收集。与 PCR 识别了， thymidine kinase 基因成功地被构造进 Ad-GFP/tk.Conclusion 包含 thymidine kinase 的复制有缺陷者侵入人体气管粘膜的病菌能被同质的再结合比常规技术在细菌更容易构造。

英文摘要：

Background Suicide gene therapy is a widely used molecular treatment for head and neck cancer. In this study, we try to use the method of homogenous recombination in bacteria to clone thymidine kinase gene （tk）-a kind of suicide gene to adenovirus backbone vectors for the construction of replication-defective adenoviruses. Methods pAdTrack-CMV/tk was constructed through subclone of a restriction endonuclease fragment including thymidine kinase gene from plasmid pCMV-tk to another plasmid pAdTrack-CMV, and then co-transfected with supercoiled pAdEasy-1, which was an adenoviral backbone vector except for deletions of E1 and E3, to competent E. coli BJ5183 for homogenous recombination using electroporation procedure. With the same method, pAdTrack-CMV was also co-transformed with pAdEasy-1 for homogenous recombination in BJ5183. Identified with restriction endonuclease Pad and polymerase chain reaction （PCR）, plasmids pAd-GFP/tk and pAd-GFP were successfully constructed. Each of them was digested with Pacl and sequently transfected into human embryo kidney 293 cells （HEK293） using Lipofectamine 2000. Results Comet-like adenovirus-producing foci of Ad-GFP/tk and Ad-GFP were observed after 5 to 7 days of cell culture After twelve days of packaging, the replication-defective adenoviruses were collected. Identified with PCR, thymidine kinase gene was successfully constructed into Ad-GFP/tk. Conclusion The replication-defective adenoviruses containing thymidine kinase can be constructed more easily by homogenous recombination in bacteria than conventional techniques.