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| 含人基质金属蛋白酶组织抑制因子-1重组腺病毒的构建与体外表达 | |
| 引用本文： | 夏冬,严律南,徐亮,童煜,左怀全,赵兰英. 含人基质金属蛋白酶组织抑制因子-1重组腺病毒的构建与体外表达[J]. 生物医学工程学杂志, 2007, 24(2): 420-424 |
| 作者姓名： | 夏冬 严律南 徐亮 童煜 左怀全 赵兰英 |
| 作者单位： | 1. 四川大学,华西医院,普外Ⅱ科,成都,610041;四川泸州医学院附属医院普外科,泸州,646000 2. 四川大学,华西医院,普外Ⅱ科,成都,610041 3. 四川泸州医学院附属医院普外科,泸州,646000 4. 成都地奥制药集团新药部,成都,610041 |
| 摘 要： | 从人肝癌组织中提取总RNA，RT—PCR合成hTIMP-1的全长cDNA，克隆到腺病毒载体AdEasy系统的穿梭质粒pAdTraek—CMV上，与骨架质粒pAdEasy-1在BJ5183受体菌中进行同源重组，成功构建含hTIMP-1全长eDNA的重组腺病毒载体，经293细胞的包装、扩增，生成含hTIMP-1基因的重组腺病毒AdhTIMP-1并实现体外表达，为进一步研究肝癌浸润和转移机理以及肝癌的基因治疗提供实验基础。 |
| 关 键 词： | 基质金属蛋白酶组织抑制因子 肝细胞癌 重组腺病毒 基因治疗 |
| 修稿时间： | 2004-11-252005-11-08 |
Construction of Recombinant Adenovirus Vector Carrying Human TIMP-1 cDNA and Its Expression In Vitro |
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| Xia Dong,Yan Lünan,Xu Liang,Tong Yu,Zuo Huaiquan,Zhao Lanying. Construction of Recombinant Adenovirus Vector Carrying Human TIMP-1 cDNA and Its Expression In Vitro[J]. Journal of biomedical engineering, 2007, 24(2): 420-424 | |
| Authors: | Xia Dong Yan Lünan Xu Liang Tong Yu Zuo Huaiquan Zhao Lanying |
| Affiliation: | 1.Department of General Surgery, West China Hospital of Sichuan University, Chengdu 610041, China; 2.Department of General Surgery, Affiliated Hospital of Luzhou Medical College 646000, China; 3.Genetic Engineering Laboratory, Chengdu Diao Group Co. Ltd, Chengdu 610041, China |
| Abstract: | The full-length cDNA of hTIMP-1 was obtained from a surgical patient with HCC by the method of RT-PCR. Then it was cloned into the adenoviral shuttle plasmid pAdTrack-CMV, and subsequently cotransformed into competent BJ5183 cells with the adenoviral backbone plasmid pAdEasy-1. Thereupon, a recombinant adenoviral plasmid containing full-length cDNA of hTIMP-1 was generated by homologous recombination in E. coli. The adenoviruses (AdhTIMP-1) were packaged and amplified in adenoviral packaging cells HEK 293. Then the viral titer was checked by green fluorescent protein (GFP), and the expression of hTIMP-1 was detected by the techniques of Western blot and RT-PCR. The recombinant adenovirus vector carrying human TIMP-1 was successfully constructed and expressed in vitro and may pave the way for further application in liver gene therapy. |
| Keywords: | Tissue inhibitor of metalloproteinases (TIMPs) Hepatocellular carcinoma (HCC) Recombinant adenovirus Gene therapy |
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