绿色荧光蛋白基因与猪瘟兔化弱毒疫苗株E2基因在杆状病毒中的融合表达

Fusion Expression of GFP and HCLV E2 Gene in Baculovirus System

为了研究猪瘟病毒E 2糖蛋白的立体结构及生物学特性,将增强型荧光蛋白基因(EGFP)和猪瘟兔化弱毒疫苗株(HCLV)E 2基因经PCR扩增后克隆至pB lueB acH is2A质粒,与杆状病毒DNA共转染后经PCR鉴定获得了含有EGFP和HCLV E 2融合基因(GFPTE 2)的重组杆状病毒rBACTE 2-339,并将其感染sf9细胞后在荧光显微镜下观察到了亮绿色荧光,说明融合基因已初步表达。

The envelope glycoprotein E2 of classical swine fever virus（CSFV） was a protective antigen inducing neutral antibody. In order to get soluble recombinant E2 glycoprotein and inspect the protein expression dynamics convinently, the en hanced green fluorescence protein（EGFP） and hog cholera lapinised virus （HCLV） E2 gene were cloned into baculovirus transfer vector pBlueBacHis2A and the recombinant plasmid pBGFPTE2 was constructed. After co-transfection pBGFPTE2 and linear baculovirus Bac-N-Blue into sf9 cell with cationic lipid reagent Cellfectin,homological recombination and plaque purification,the pure recombinant baculovirus clone named rBACTE2-339 were harvested and proliferated in sf9 cell to generate P2 stock. After infection with P2 stock,brilliant green fluorescence was detected under the fluorescence microscopy confirming the heterogenous protein expression of recombinant baculovirus in sf9 cell.