Construction of Recombinant DNA Plasmid Co-expressing Capsid Protein of Porcine circovirus Type 2 and Porcine Granulocyte-macrophage Colony-stimulating Factor and Its Expression In vitro
SONG Qin-ye;YANG Han-chun;GUO Xin;CHEN Yan-hong;CHA Zhen-lin
Key Laboratory for Preventive Veterinary Medicine, Ministry of Agriculture, China Agricultural University, Beijing 100094, China
摘要将猪圆环病毒2型(Porcine circovirus type 2, PCV2)orf2和猪粒细胞-巨噬细胞集落刺激因子(gm-csf)基因插入同一哺乳动物细胞表达载体pIRESneo中，构建出重组共表达质粒pIRES-orf2/gm-csf。以脂质体转染方法将重组质粒pIRES-orf2/gm-csf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orf2和gm-csf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-orf2/gm-csf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。
Abstract：The orf2 gene of Porcine circovirus type 2 (PCV2) and porcine granulocyte-macrophage colony-stimulating factor (gm-csf) were inserted into pIRESneo to construct recombinant plasmid pIRES-orf2/gm-csf. The recombinant plasmid pIRES-orf2/gm-csf was then transfected transiently into COS7 cells, the expressions of orf2 and gm-csf genes were confirmed by indrect immunofluorscence assay (IFA) and colony-stimulation forming assay. The results showed that the transfected cells could display specific immunofluorscence and the supernatant of cell culture had the activity for stimulating the formation of porcine bone marrow cell colonies. The successful construction of pIRES-orf2/gm-csf co-expressing PCV2 orf2 and gm-csf genes provides foundation for the further study of the PCV2 DNA vaccine.
宋勤叶;杨汉春;郭鑫;陈艳红;查振林. 共表达PCV2衣壳蛋白和猪GM-CSF重组质粒的构建与体外表达[J]. , 2006, 14(3): 307-311.
SONG Qin-ye;YANG Han-chun;GUO Xin;CHEN Yan-hong;CHA Zhen-lin. Construction of Recombinant DNA Plasmid Co-expressing Capsid Protein of Porcine circovirus Type 2 and Porcine Granulocyte-macrophage Colony-stimulating Factor and Its Expression In vitro. , 2006, 14(3): 307-311.
