Computational modeling and functional analysis of Herpes simplez virus type-1 thymidine kinase and Escherichia coli cytosine deaminase fusion protein
Herpes simplex virus type-1 thymidine kinase (HSV-1TK) and Escherichia coli cytosine deaminase (CD) fusion protein was designed using Insightll software. The structural rationality of the fusion proteins incorporating a series of flexible linker peptide was analyzed, and a suitable linker peptide was chosen for further investigated. The recombinant plasmid containing the coding regions of HSV-1TK and CD cDNA connected by this linker peptide coding sequence was generated and subsequently transfected into the human embryonic kidney 293 cells (HEK.293). The Western blotting indicated that the recombinant fusion protein existed as a dimer with a molecular weight of approximately 90 kDa. The toxicity of the prodrug on the recombinant plasmid-transfected human lung cancer cell line NCIH460 was evaluated, which showed that TKglyCD-expressing cells conferred upon cells prodrug sensitivities equivalent to that observed for each enzyme independently. Most noteworthy, cytotoxicity could be enhanced by concurrently treating TKglyCD-express ing cells with prodrugs GCV and 5-FC. The results indicate that we have successfully constructed a HSV-lTKglyCD fusion gene which might have a potential application for cancer gene therapy.
Molecular modeling Fusion peptide Cytosine deaminase Herpes simplez virus type-1 thymidine kinase
Jufeng Zhang Zhanli Wang Fang Wei Wei Qiu Liangren Zhang Qian Huang
Central Experimental Laboratory, the First Peoples Hospital, Shanghai Jiaotong University, Shanghai Technology Center, NeoTrident Technology Ltd., Beijing 100080, China School of Pharmaceutical Science, Peking University, Beijing 100083, China
国际会议
广州
英文
322-326
2008-11-01（万方平台首次上网日期，不代表论文的发表时间）