【摘要】 根据GenBank上WSSV囊膜蛋白基因vp28的序列,设计合成引物,PCR扩增得到vp28基因,成功构建重组表达载体pBAD/gⅡIA-VP28并转化大肠杆菌E.coli。用L-阿拉伯糖在37℃诱导重组基因工程菌,表达产物经SDS-PAGE和Western-blot检测,显示有与预期大小31kDa相符合的目的蛋白。荧光显微镜方法分析显示,表达的VP28可与克氏原螯虾血细胞结合。结果表明,在合适的培养条件下,构建的重组表达载体pBAD/gⅡIA-VP28不仅能够表达vp28基因,而且表达的VP28具有很高的抗原结合活性。更多还原

【Abstract】 In this study,a pair of primers was designed by primer design software 5.0 according to the sequence of vp28 gene of white spot syndrome virus (WSSV) in GenBank. The vp28 DNA fragment was amplified by PCR and cloned into Escherichia coli (E.coli) expression vector pBAD/gⅢA successfully. The pBAD/gⅢA-VP28 was then transformed into E.coli.After L-arabinose induction at 37℃,the fusion protein with 31 kDa was expressed,which was confirmed by SDS-PAGE and Western-blot analysis. Binding assay by fluorescence microscopy in vitro showed that VP28 was capable of binding to shrimp hemocytes. Results confirmed that the recombinant vector pBAD/gⅢA-VP28 can express vp28 gene of WSSV and the VP28 had high antigenic activity.更多还原