猪圆环病毒Ⅱ型贵州分离株ORF2基因表达的试验  

Study for Expression of Porcine circovirus Guizhou Strain ORF2 Gene

作　　者：韦冠东[1] 甘振磊[1] 马萍[2] 汤德元[1] 王洪光[1] 唐宇[2] 张元鑫[2] 曾智勇[1] 李达[1] 

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025 [2]贵州省动物疫病预防控制中心,贵州贵阳550008

出　　处：《中国兽医杂志》2015年第9期43-46,共4页Chinese Journal of Veterinary Medicine

基　　金：贵州省科学技术基金项目资助[黔科合J字(2013)2110号];贵州省2011年农业攻关项目资助[黔科合NY字(2011)3103号];贵州省2010年农业科技攻关项目[黔科合NY字(2010)3042号]

摘　　要：通过采集疑似猪圆环病毒感染的病料,分离到1株猪圆环病毒2型贵州株,并对其进行了鉴定。原核表达试验结果表明,PCV-2贵州株的ORF2基因可在大肠杆菌BL21(DE3)中表达,表达的目的蛋白约为40.0 k Da,且复性蛋白具有一定免疫原性。将真核表达质粒分别转染PK-15细胞,以掌握ORF2基因和白细胞介素-2(IL-2)基因体外表达情况;将重组质粒、空载体及生理盐水分别免疫BALB/c小鼠,以监测血清中特异性抗体及T淋巴细胞亚群含量,从而探讨猪圆环病毒Ⅱ型贵州分离株ORF2基因真核表达效果及长白猪细胞因子IL-2对PCV2 ORF2免疫原性的影响,结果表明,pc DNA3.1-ORF2和pc DNA3.1-ORF2-IL-2在PK-15细胞中获得了较高的表达,IL-2可明显增强PCV-2 DNA免疫效果。A porcine circovirus type 2 virus strain was isolated and identified in samples from infected pigs in this study.The result of prokaryotic expression showed that the ORF2 gene could express in E.coli BL21（DE3）,and the expressed protein was about 40 KD with a certain degree of immunogenicity. To study the eukaryotic expression of porcine circovirus Guizhou Strain ORF2 gene and the effect of pig cytokine IL-2 on PCV2 ORF2 immunogenicity,eukaryotic expression plasmids were transfected into PK-15 cells to study the expression of ORF2 gene and IL-2 gene in vitro.And then BALB/c mice were immunized with recombinant plasmids empty vector and saline and the specific serum antibodies and T lymphocyte subsets,were analyzed. Our results showed that pc DNA3.1-ORF2 and pc DNA3.1-ORF2-IL-2 were expressed efficiently in PK-15 cells,and IL-2 could significantly enhance the immune response of PCV2 DNA.

关 键 词：猪圆环病毒2型 ORF2基因 IL-2 表达 重组质粒 

分 类 号：S852.651[农业科学—基础兽医学]