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目的研究L612K变异MxA蛋白抑制水泡性口膜炎病毒(VSV)复制活性。方法将野生型MxA蛋白重组表达载体pcDNA3.1-MxA(WT)和L612K变异MxA蛋白载体pcDNA3.1-MxA(L612K)分别瞬时转染Wish细胞,24h后VSV感染细胞,感染后48hMTT检测各组细胞增殖数;另取Wish细胞转染MxA载体和对照DNA,24h后加入VSV,感染24h后收集细胞,RT-PCR检测VSV mRNA水平,Western blot检测MxA蛋白的表达水平。结果野生型和L612K变异MxA蛋白均在Wish细胞有较好表达;MTT检测L612K变异组细胞增殖数明显低于野生型(t=1.13,P<0.01),RT-PCR检测L612K变异组VSV mRNA水平明显高于野生型组(t=0.13,P<0.01)。结论L612K变异可能使MxA蛋白降低了对VSV的抑制活性。
Objective To investigate the inhibitory effect of L612K variant MxA on vesicular stomatitis virus (VSV) replication. Methods Transfect Wish cells with wild-type MxA recombinant expression vector pcDNA3.1-MxA (WT) and L612K mutant MxA protein vector pcDNA3.1-MxA (L612K) transiently. After VSV infection for 24 hours, Cells were transfected with Wish cells. VSV was added after 24 hours and cells were collected 24h after infection. VSV mRNA levels were detected by RT-PCR and MxA protein was detected by Western blot. Results Both wild-type and L612K mutant MxA proteins were expressed well in Wish cells. The proliferation of L612K mutation group was significantly lower than that of wild-type strain by MTT assay (t = 1.13, P <0.01) Significantly higher than the wild-type group (t = 0.13, P <0.01). Conclusion L612K mutation may reduce MxA protein’s inhibitory activity on VSV.