轮状病毒VP3表达载体pEGFP-VP3的构建、表达及功能研究

Construction,expression and function study of rotavirus VP3 expression vector pEGPF-VP3

通过构建重组表达载体pEGFP-VP3,将pEGFP-VP3转染MA104细胞,观察VP3的表达情况和表达的VP3对轮状病毒复制的作用,进一步评价构建方式是否成功。GFP荧光和Western Blot结果显示,pEGFP-VP3转染MA104细胞48 h,重组蛋白VP3表达量最高。通过RV拷贝数和免疫荧光检测病毒的复制情况,结果均显示重组蛋白VP3可促进病毒复制,为后续更加深入地探究VP3的功能及在轮状病毒复制中的作用提供了实验基础。

The VP3 gene was inserted into the eukaryotic expression vector pEGPF-N2,and further transfected the recombinant plasmid into MA104 cells to observe whether the expressed VP3 has an effect on rotavirus replication.The expression of recombinant protein was detected by immunofluorescence and Western Blot.The expression of VP3 was the strongest at 48 h post transfection in MA104 cells.And the replication of the virus was detected by qRT-PCR and immunofluorescence.The results showed that the recombinant protein VP3 could promote viral replication,which provided an experimental basis for further exploration of the function of VP3 and its role in rotavirus replication.