猪2型圆环病毒核酸疫苗的构建、鉴定和表达

Construction ,Identification and Expression of PCV2 Nucleotide Vaccines

以pVAX1为真核表达载体,以猪白介素18基因、猪2型圆环病毒内蒙古株的衣壳蛋白和复制蛋白基因为目的基因,构建了pVAX1-ORF2、pVAX1-IL18-ORF2、pVAX1-ORF2-ORF1和pVAX1-IL18-ORF2-ORF1等4个重组质粒作为核酸疫苗。将这4个重组核酸质粒分别酶切鉴定后,转染BHK-21和PK-15细胞进行RT-PCR、间接免疫荧光和Western blotting检测。结果表明,重组核酸质粒构建正确,并能进行表达。

Using pVAX1 as eukaryotic expressing vector,swine interkine 18 genes,PCV2（Innermongolia strain） capsid and replicated protein genes as aimed genes, 4 recombinant plasmids were contructed as nucleotide vaccines. They are named as pVAX1-ORF2, pVAX1-ORF2-ORF1, pVAX1-IL18-ORF2, pVAXI-IL18-ORF2-ORF1. The four recombinant plasmids were transfected into BHK-21 cell after enzyme-identificated,and detected by RT-PCR,indirect immunofluorescence and Western blotting. The result shows that the nucleotide vaccines were constructed correctly and expressed in cells.