变形链球菌葡聚糖结合蛋白A真核表达质粒pcDNA3.1/GBD在哺乳动物细胞中的表达
Expression of GBD Gene ofStreptococcus MutansGlucan Binding Protein A in Mammalian Cells
作者：苏凌云1,吴补领1,李福洋2,陆 群1
Author：SULing-yun1,WUBu- ling1,LI Fu-yang2,LUQun1
收稿日期：2004-02-25 年卷（期）页码：2004,22(01):10-
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：变形链球菌,葡聚糖结合蛋白,哺乳动物细胞,基因表达,
Key words：Streptococcus mutans,glucan-binding protein,mammalian cells,gene expression,
基金项目：高等学校骨干教师资助计划资助项目(2000年)
中文摘要
目的 观察变形链球菌葡聚糖结合蛋白A(GbpA)的葡聚糖结合区(GBD)真核表达质粒pcDNA3·1/GBD在哺乳动物细胞COS-7的表达情况。方法 通过基因重组技术构建真核表达质粒pcDNA3·1/GBD,通过脂质体转染法将其转染至COS-7细胞,通过免疫组化SABC法检测其在COS-7细胞的表达。结果 pcDAN3·1/GBD质粒转染的细胞质呈棕黄色染色,胞核无着色,pcDAN3·1空载体转染的细胞质及胞核无着色,空白对照组也无着色。结论 质粒 pcDAN3·1/GBD转染COS-7后能够在细胞内翻译、表达,表达的蛋白质位于细胞质中,可与抗GbpA抗体特异性结合,具有抗原性,可作为基因疫苗。
英文摘要
Objective To evaluate the expression of recombinant plasmid pcDNA3·1/GBD of glucan binding protein ofStrepto- coccus mutansin mammalian cells COS-7.Methods Eukaryotic plasmid carrying encoding gene of GBD ofStreptococcus mutans gbpAwas constructed and the plasmid was introduced into COS-7 cells by Lipofectamine reagent. The transient expressed protein in COS-7 cells was detected by immunochemistry technique.Results The positive expression was detected in plasma of the cells which were transfected with recombinant plasmid pcDNA3·1/GBD. The cells which were transfected with pcDNA3·1 were nega- tive.Conclusion GBD can translate and express in COS-7 cells aftertransfectedwith recombinant plasmid pcDNA3·1/GBD. The expressed protein locates in the plasma and the protein is able to combine with anti-GbpA antibody. The expressed protein has the antigenicity and is a candidate gene vaccine.
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