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目的研究T103A变异MxA蛋白抑制水疱性口膜炎病毒(VSV)复制活性。方法将野生型、T103A变异MxA蛋白表达载体和对照质粒分别瞬时转染Wish细胞,24h后VSV感染细胞,48h后用MTT法检测各组细胞增殖;另取Wish细胞转染上述3种质粒,转染24h加入VSV感染细胞,24h后收集细胞采用RT-PCR检测VSV mRNA水平;Western blot检测各组MxA蛋白表达。结果野生型、T103A变异MxA蛋白均在Wish细胞有较好表达;MTT检测结果提示T103A变异组细胞增殖数显著低于野生型组(P<0.01);RT-PCR结果显示T103A变异组VSVmRNA水平显著高于野生型组(P<0.01),但与对照组比较差异无统计学意义(P>0.05)。结论T103A变异MxA蛋白失去了抑制VSV复制活性。
Objective To study the inhibitory effect of T103A mutant MxA protein on the replication activity of vesicular stomatitis virus (VSV). Methods The wild-type and T103A mutant MxA protein expression vectors and control plasmids were transiently transfected into Wish cells, VSV infected cells after 24h, 48h after MTT assay of the proliferation of each group; another Wish cells transfected the above three plasmids, transfer The cells were infected with VSV for 24 h and VSV mRNA was detected by RT-PCR after 24 h. Western blot was used to detect the expression of MxA in each group. Results The wild-type and T103A mutant MxA proteins were expressed well in Wish cells. The results of MTT assay showed that the proliferation of T103A mutant group was significantly lower than that of wild-type group (P <0.01). The RT-PCR results showed that the level of VSV mRNA in T103A mutation group was significantly (P <0.01), but there was no significant difference compared with the control group (P> 0.05). Conclusion T103A mutant MxA protein has no inhibitory activity on VSV replication.