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为了获得具有良好免疫学活性的猪细小病毒(PPV)基因工程抗原,试验采用原核表达方法对猪细小病毒NS1蛋白进行了表达研究。结果表明:成功克隆了大小为1 389 bp的NS1基因主要抗原区域,经IPTG诱导获得了大小为60 ku、以包涵体形式表达的重组蛋白。利用6×His-Tagged Protein Purification Kit获得了高纯度的纯化蛋白,纯化蛋白能与PPV阳性血清发生特异性反应,以纯化蛋白作为包被抗原的间接ELISA方法与血凝抑制试验(HI)的符合率为97.92%。说明试验成功获得了体外表达的NS1重组蛋白,且重组蛋白具有良好的免疫学活性。
In order to obtain porcine parvovirus (PPV) genetically engineered antigen with good immunological activity, the prokaryotic expression method was used to study the expression of porcine parvovirus NS1 protein. The results showed that the major antigenic region of NS1 gene with the size of 1 389 bp was successfully cloned and the recombinant protein was expressed in the form of inclusion body with the size of 60 ku induced by IPTG. The purified protein was purified with 6 × His-Tagged Protein Purification Kit. The purified protein reacted specifically with PPV-positive sera. The indirect ELISA method using the purified protein as coating antigen was in agreement with the hemagglutination inhibition test (HI) The rate was 97.92%. This indicated that the NS1 recombinant protein expressed in vitro was successfully obtained and the recombinant protein had good immunological activity.