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从对虾白斑综合症病毒基因组的预测开放阅读框中选取3个编码类锌指蛋白的基因wsv063、wsv069和wsv477,将它们克隆到pET-GST表达载体,以E.coliBL21(DE3)为宿主菌,成功表达了GST融合的目的蛋白.表达的GST-WSV063和GST-WSV477在菌体里主要以包涵体形式存在,GST-WSV069在菌体里可溶性形式和包涵体形式几乎等量存在.采用glutathione sepharose 4B或Ni-NTA agarose亲和层析对目的蛋白进行了纯化.以上3种病毒蛋白的表达和纯化为进一步研究它们的功能奠定了基础.
Three genes coding for zinc finger protein wsv063, wsv069 and wsv477 were selected from the predicted open reading frame of WSSV genome and cloned into pET-GST expression vector. E.coli BL21 (DE3) GST-WSV063 and GST-WSV477 were expressed mainly in the form of inclusion bodies in GST-WSV069 cells, while GST-WSV069 was almost equal in soluble forms and inclusion bodies in G-cells.Using glutathione sepharose 4B or Ni-NTA agarose affinity chromatography.The expression and purification of the above three viral proteins laid the foundation for further study of their function.