登革病毒prM基因真核表达载体的构建及在白纹伊蚊细胞C6/36中的表达

Construction of eukaryotic expression vector containing prM gene from dengue virus and expression in Aedes albpictus cells C6/36

目的 构建登革病毒prM基因重组表达质粒并在白纹伊蚊细胞C6 / 36中表达。方法 将prM基因亚克隆入真核表达载体 pEGFP -N3,转化大肠杆菌JM 10 9,筛选阳性克隆进行PCR及酶切鉴定 ,将获得的重组质粒以脂质体法转化白纹伊蚊C6 / 36细胞并使其表达 ,利用RT -PCR法检测目的基因的转录 ,Western -blot检测融合蛋白的表达。结果 成功构建了 pEGFP -prM重组质粒 ,RT -PCR证明 prM基因在蚊细胞内的转录 ,Western -blot检测到 prM -GFP融合蛋白的表达。结论 成功构建登革了病毒 prM基因重组表达质粒并在白纹伊蚊细胞C6 / 36中表达 。

Aim To construct recombinant plasmid containing prM gene from dengue virus and make it to express in Aedes albopictus cells C6/36 Methods prM gene was subcloned into pEGFP N3 vector The recombinant plasmid was transformed into E coli JM109 Recombinant clone was identified by PCR and endonuclease digestion The recombinant plasmid was transfected into C6/36 cells The expression product was identified by RT PCR and Western blot Results prM GFP recombinant plasmid has been successfully constructed RT PCR and Western blot identified the expression of prM gene in mosquito cells C6/36 Conclusion prM GFP recombinant plasmid has been successfully constructed The prM GFP fusing protein is expressed in mosquito cells C6/36 It can be used in further study for the intracellsular immunization and transgenic mosquito