共表达PCV2衣壳蛋白和猪GM-CSF重组质粒构建和体外表达.pdf

业生物技术学报 Journal of Agricultural Biotechnology 2006,14 (3): 307~3 11 ·研究论文· 共表达PCV2 衣壳蛋白和猪GM-CSF 重组质粒的构建与体外表达* 勤叶, 杨汉春**, 郭 鑫, 陈艳红, 查振林 (中国农业 学 业部预防兽医学重点开放实验室，北京 100094) ： type 2, PCV2) 和猪粒细胞- 巨噬细胞集落刺激因子( )基因插入同一 摘要 将猪圆环病毒2 型( 哺乳动物细胞表达载体 pIRESneo 中，构建出重组共表达质粒 pIRES- 。以脂质体转染方法将重组质粒 pIRES- 转染COS7 细胞，经间接免疫荧光实验和集落刺激形成实验检测，结果表明 和 基因在COS7 细 胞中得到表达，转染细胞呈现特异性荧光，转染细胞培养上清能剌激猪骨髓细胞形成集落。重组质粒pIRES- 的成功 构建为进一步研究猪圆环病毒2 型的DNA 疫苗奠定了基础。 ： ；猪粒细胞- 巨噬细胞集落刺激因子；重组质粒；共表达 关键词 猪圆环病毒2 型； 中图分类号: S188 文献标识码: A 文章编号: 1006-1304(2006)03-0307-05 Construction of Recombinant DNA Plasmid Co-expressing Capsid Protein of Type 2 and Porcine Granulocyte-macrophage Colony-stimulating Factor and Its Expression SONG Qin-ye, YANG Han-chun**, GUO Xin, CHEN Yan-hong, CHA Zhen-lin The gene of type 2 (PCV2) and porcine granulocyte-macrophage colony-stimulating factor ( ) were inserted into pIRESneo to construct recombinant plasmid pIRES- . The recombinant plasmid pIRES was then transfected transiently into COS7 cells, the expressions of and genes were confirmed by in- drect immunofluorscence assay (IFA) and colony-stimulation forming assay. The results showed that the transfected cells could dis- play specific immunofluorscence and the supernatant of cell culture had the activity for stimulating the formation of porcine bone mar- row cell colonies. The successful construction o