摘要
目的:构建变形链球菌表面蛋白A区(spap/A)真核表达质粒pVAX1-spap/A,并观察其在哺乳动物细胞COS-7中的表达。方法:通过基因重组技术,构建真核表达质粒pVAX1-spap/A。采用脂质体转染法,将其转染至COS-7细胞中,然后经免疫组化SABC法检测其在细胞中的表达。结果:真核表达质粒pVAX1-spap/A经酶切分析,证实携带spap/A片段。pVAX1-spap/A转染的细胞胞质呈褐色,pVAX1空质粒转染的细胞胞质中无着色。结论:成功构建真核表达质粒pVAX1-spap/A,所携带的基因序列正确,能够在真核细胞COS-7中正确表达目的蛋白,为下一步基因防龋疫苗的研究奠定了基础。
PURPOSE： The study is aimed to evaluate the expression of recombinant plasmid pVAXl-spap/A of surface protein antigen A of Streptococcus mutans in mammalian cells COS-7. METHODS： The eukaryotic plasmid carrying encoding gene of spap/A of Streptococcus mutans was constructed and the plasmid introduced into COS-7 cells by lipofeetamine reagent. The transient expressed protein was detected by immunochemistry technique in COS-7 cells. RESULTS： Positive expression was detected in plasma of the cells which were transfected with recombinant plasmid pVAX1-spap/A. The cells which were transfected with pVAX1 were negative. CONCLUSIONS： Spap/A can translate and express in COS-7 cells after transfected with recombinant plasmid pVAX1-spap/A. The expressed protein locates in the plasma and the protein is able to combine with anti-spap/A antibody. The expressed protein has the antigenicity and recombinant plasmid pVAXl-spap/A is a candidate vaccine.
出处
《上海口腔医学》
CAS
CSCD
2009年第1期61-65,共5页
Shanghai Journal of Stomatology
基金
贵州省教育厅重点项目(黔教科2004119)~~
关键词
变形链球菌
表面蛋白
PVAX1
真核表达
Streptococcus mutans
Surface protein antigen
pVAX1
Eukaryotic expression
