猪圆环病毒2型重组病毒ZJ-R感染性克隆的构建

Construction of an infectious clone of porcine circovirus type 2 recombinant virus ZJ-R

利用无缝克隆方法构建ZJ-R全长基因组的单拷贝分子克隆以及双拷贝串联分子克隆;通过生物信息学技术预测ZJ-R病毒结构蛋白的B细胞抗原表位,人工合成选定的表位肽,然后偶联KLH免疫新西兰大白兔制备多克隆抗体;免疫组化试验证实该多抗与转染PK15细胞的ZJ-R双拷贝串联分子克隆具有反应性。结果表明构建的ZJ-R双拷贝串联分子克隆具有感染性。

The research was conducted to construct an infectious clone of porcine circovirus type 2 recombinant virus ZJ-R. One copy and two copies in tandem of the whole genomic sequence of ZJ-R were cloned into the pSK vector with restriction site of enzyme EcoRV. The B cell epitopes of the ZJ-R virus structural protein were predicted by using biotic soft-wares. The epitope peptides were chosen, synthesized and coupled to KLH. Then the New Zealand White Rabbits were immunized to elicite the polyclonal antibodies, which were used later in immunohistochemical techniques. The results showed that the ZJ-R virus could be detected by IHC after 72 h post transfection of pSK-2 ZJ-R. In conclusion, an infectious DNA clone of the porcine circovirus type 2 recombinant virus ZJ-R was constructed suc- cessfully.