pVVP3和pVHN核酸疫苗的构建、表达及对肿瘤细胞的作用  

Construction and Expression of the Nucleic Acid Vaccine pVVP3 and pVHN and Its Effect on Tumor Cell

作　　者：米志强[1] 金宁一[1] 龚伟[1] 薛立娟[1] 连海[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：《中国生物化学与分子生物学报》2003年第6期704-708,共5页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金重大项目 (No .3 9893 2 90 4 3 );国家"973"项目(No .G1990 1190 2 )资助~~

摘　　要：用pVAX1载体构建抗肿瘤核酸疫苗 .将鸡贫血病病毒 (CAV)的VP3基因和鸡新城疫病毒(NDV)HN基因插入载体pVAX1的多克隆位点 ,构建了 2个重组基因疫苗pVVP3和pVHN .转染OS732细胞 ,Western印迹及血凝试验检测HN基因表达状况及表达产物的活性 .RT PCR、DNALadder、TUNNEL染色检测VP3基因的表达及表达产物诱导OS732细胞凋亡作用 .酶切鉴定证实成功地构建了两个核酸疫苗 ,并能在真核细胞内表达 .含HN的核酸疫苗pVHN表达后具有血凝活性并致肿瘤细胞表面唾液酸含量降低 (P <0 0 5 ) .pVVP3的转染能导致OS732细胞凋亡 ,凋亡率可达87% .试验结果表明 。Nucleic acid vaccine pVVP3 and pVHN were constructed with HN gene from NDV and VP3 gene from CAV, respectively, using pVAX1 vector. pVVP3 and pVHN were transfected into OS732 tumor cell. HN gene expression was detected by Western blotting and hemagglutination test. The content of OS732 cell surface sialic acid was measured. The influence of VP3 gene on OS732 was investigated by TUNNEL assay and DNA Ladder electrophoresis. Enzyme restriction test indicated that pVVP3 and pVHN were correctly constructed. Apoptosis was induced by pVVP3 on OS732 tumor cell with the rate of 87%. The HN expression could reduce the sialic acid content of OS732. The result indicated that the nucleic acid vaccine pVVP3 and pVHN had potential antitumor effect.

关 键 词：鸡新城疫病毒 HN基因 鸡贫血病病毒VP3基因 抗肿瘤 

分 类 号：S852.65[农业科学—基础兽医学] S858.31[农业科学—兽医学]