共表达VP3与hIL-18基因真核重组质粒的构建及对人结肠癌细胞的影响  

Construction and Expression of Eukaryotic Recombinant Plasmid Including CAV VP3 Gene and Human IL-18 Gene and Influence on Colon Cancer Cell

作　　者：金宁一[1] 连海[1] 米志强[1] 李肖[1] 徐敬龙[1] 金洪涛[1] 解丽华[1] 李萍[1] 

机构地区：[1]解放军军需大学解放军基因工程重点实验室,长春130062

出　　处：《高技术通讯》2003年第9期43-46,共4页Chinese High Technology Letters

基　　金：973规划(G1999011902)资助项目。

摘　　要：先将鸡贫血病毒VP3基因插入真核表达载体pVAX1的多克隆位点,再将pIRES1载体上的IRES启动子及新霉素基因一同切下插入pVAX1的VP3基因下游,然后切下新霉素基因,将hIL-18基因插入其中,构建成pVVP3IL-18,将pVVP3IL-18转染Hela细胞。间接免疫荧光表明,在细胞膜和细胞浆观察到了特异的黄绿色荧光,证明表达了hIL-18。pVVP3IL-18转染人结肠癌细胞HCT-116后,经AO/EB染色及电镜观察,可见大量细胞凋亡。说明真核重组质粒pVVP3IL-18可在HeLa细胞中表达,并可在体外诱导人结肠癌细胞HCT-116凋亡。IRES promotor and neomycin gene from pIRES1 vector were inserted into multiple clone site of pVAX1 vector, then a recombinant plasmid pWP3IL-18 was constructed by inserting CAV VP3 gene and human IL-18 gene into multiple clone site and neomycin site , respectively. The recombinant plasmid pVVP3IL-18 was transfected to Hela cells and the results of indirect immunofluorescence showed that specific bright yellow green flurescene was observed around the membrane and across the plasma of the transfected Hela cells when tested with anti-mouse Ig G, FITC conjugate. The results of AO/EB staining and electron microscope showed apoptosis was induced greatly after pVVP3IL-18 was transfected to HCT-116 cells. The above results showed that the recombinant plasmid pVVP3IL-18 can express in Hela cells and can induce apoptosis in HCT-116 cells.

关 键 词：鸡贫血病毒 真核表达 载体 VP3 hIL-18 IRES启动子 HELA细胞 结肠癌细胞 凋亡素 真核重组质粒 

分 类 号：R735.35[医药卫生—肿瘤]