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中文摘要：
猪博卡病毒是近年来新发现的一种细小病毒。本研究根据其部分基因组序列（GenBank登录号GU902967-GU902971）设计了一对引物,采用PCR方法扩增出了猪博卡病毒的NP1基因,将其克隆到表达载体pET32a（＋）中,构建了重组质粒pET32a-NP1,经测序鉴定正确后,将其转化感受态细胞BL21（DE3）中,并进行IPTG诱导表达。结果表明,重组菌可表达分子量为46 kDa的融合蛋白,蛋白表达量较高,而且蛋白以可溶性形式存在于菌体中。表达产物经纯化后,目的蛋白纯度较好。Western Blot结果显示,表达的NP1蛋白能与His抗体发生特异性的免疫反应,表明原核表达的NP1蛋白具有良好的反应原性。
英文摘要：
Porcine bocavirus（PBoV） was a recently discovered parvovirus from pigs.One pair of primers was designed according to the published sequences of genome of the porcine bocaviruses（GenBank accession number GU902967-GU902971）.The NP1 gene was amplified by PCR and cloned into the prokaryotic expression vector pET32a（＋） to construct a recombinant pET32a-NP1.After sequencing,the recombinant was transformed into Escherichia coli BL21（DE3） competent cells.The transformed bacteria were induced by IPTG to produce a recombinant protein of 46 kDa of molecular weight.The result showed that recombinant protein was higher expression and purity by the Ni-NTA resin,and in soluble form in the bacteria.The recombinant protein could react with the antibody of His in a Western blot test,indicating that the expressed protein possessed strong reactinogenicity.
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