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目的 构建一种肝癌细胞靶向性的腺相关病毒载体以用于肝癌的促凋亡基因治疗的研究。方法 通过设计含有特定酶切位点的引物 ,选择性地从人基因组中扩增出人甲胎蛋白 (α fe toprotein ,AFP)启动子序列并克隆到真核表达载体pTR UF5上 ;再通过平端连接的方法 ,构建成含甲胎蛋白启动子和人野生型p5 3基因的重组腺相关病毒 (recombinantadeno associatedvirus,rAAV)质粒rAAV AFP 5 3和绿色荧光蛋白基因GFP(greenfluorescenceprotein)的质粒rAAV AFP GFP。结果 成功地构建了以腺相关病毒为载体、受人甲胎蛋白启动子调控表达人野生型p5 3基因的质粒系统。结论从理论上说 ,我们构建的重组腺相关病毒质粒rAAV AFP p5 3,可以特异性地转染到肝癌细胞中 ,从而为肝癌基因治疗提供一种有效手段
OBJECTIVE: To construct a hepatocellular carcinoma cell-targeted adeno-associated virus vector for hepatocellular carcinoma-related gene therapy. Methods The promoter of human α-fetoprotein (AFP) was amplified from the human genome by designing primers containing specific restriction sites and cloned into eukaryotic expression vector pTR UF5. , A recombinant rAAV AFP GFP plasmid containing the recombinant adeno associated virus (rAAV) plasmid rAAV AFP 5 3 and green fluorescent protein (GFP) containing the alpha-fetoprotein promoter and human wild-type p5 3 gene was constructed. Results A plasmid system was successfully constructed to express human wild-type p5 3 gene under the control of human alpha-fetoprotein using adeno-associated virus as a vector. Conclusions In theory, we constructed the recombinant adeno-associated virus plasmid rAAV AFP p5 3, can be specifically transfected into liver cancer cells, and thus provide an effective means for liver cancer gene therapy