【摘要】 为研发猪萨佩罗病毒(PSV)检测试剂,根据PSV YC2011毒株核苷酸序列,设计针对1D基因的特异引物,以YC2011毒株为模板,利用RT-PCR方法,成功扩增出1D基因,将该基因与原核表达载体pET-32a(+)连接,转化表达菌株BL21。经PCR和测序鉴定,阳性菌株IPTG诱导表达,融合蛋白1D进行SDS-PAGE和Western blot检测分析。结果表明,成功构建了原核表达菌株,其所表达的融合蛋白分子质量约为51ku,且可被猪萨佩罗病毒阳性血清所识别。更多还原

【Abstract】 According to the porcine Sapelovirus YC2011strain nucleotide sequence reported in the GenBank, we designed a pair of primers to amplify 1Dgene of YC2011strain.The 1Dgene was cloned into prokaryotic expression vector pET-32a(+)and transformed into E.coli strain BL21.In order to obtain 1Dprotein, the positive strains were induced by IPTG,and then the expressed products were analyzed by SDS-PAGE and Western blot.The results of SDS-PAGE and Western blot demonstrated that the 1Dgene of YC2011 strain was successfully expressed.The expressed 1Dprotein has 51ku,and can be recognized by the positive serum of Porcine sapelovirus.更多还原