猪圆环病毒Cap基因重组质粒的构建及对小鼠免疫效果研究  

Construction of Recombinant Plasmid with Porcine Circovirus Cap Gene and Its Immune Effects in Mice

作　　者：刘自立[1,2] 张志[2] 吴发兴[2] 刘爽[2] 董雅琴[2] 邵卫星[2] 段纲[1] 李晓成[2] 

机构地区：[1]云南农业大学动物科学技术学院,云南昆明650201 [2]中国动物卫生与流行病学中心,山东青岛266032

出　　处：《动物医学进展》2014年第2期37-41,共5页Progress In Veterinary Medicine

基　　金：广西科技攻关项目(桂科攻0815009-3-9);科技部科技基础性工作专项(2012FY111000)

摘　　要：用PCR方法扩增出猪圆环病毒2型的Cap基因,将1个Cap基因、2个串联Cap基因和3个串联Cap基因定向克隆于真核表达载体pcDNA4-His/Max,构建真核表达载体pcDNA4-His/Max-Cap、pcDNA4-His/Max-2Cap和pcDNA4-His/Max-3Cap。将构建好的重组质粒以100μg/只腿部肌肉注射昆明小鼠,同时设置pcDNA4-His/Max和生理盐水对照,于首免后14d、28d再次免疫,并分别于首免后7、14、28、35、49d用ELISA方法检测小鼠抗体水平,对不同串联重组质粒免疫原性进行初步评价。结果显示,重组质粒于35d能诱导小鼠产生较高的体液免疫,2个Cap和3个Cap串联基因的免疫效果远远高于1个Cap基因的免疫效果,差异极显著(P<0.01)。Cap gene of porcine circovirus type 2 was amplified by PCR and cloned into the eukaryotic expres- sion vector pcDNA4-His/Max to construct a series of recombinant vectors pcDNA4-His/Max-Cap, pcD- NA4-His/Max-2Cap, pcDNA4-His/Max-3Cap by using one Cap, two Cap and three Cap genes. The con- structed recombinant plasmid was intramuscularly injected into Kunming mice with 100 g and re-immuni- zation at 14, 28 day, meanwhile a blank plasmid and saline control were established. The antibody levels in mice were determined by ELISA at 7, 14, 28, 35, 49 day after inoculation. The results showed that re- combinant plasmid was able to induce satisfactory humoral immunity and the 3Cap recombinant vectors have high immunogenicity compared with other recombinant vector.

关 键 词：猪圆环病毒 Cap基因 核酸疫苗 

分 类 号：S852.659.2[农业科学—基础兽医学] Q789[农业科学—兽医学]