L612K变异MxA蛋白体外抑制水泡性口膜炎病毒复制的研究

Inhibition of vesicular stomatitis virus replication by MxA protein with L612K mutant in vitro

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 antiviral activity of L612K mutant MxA protein against vesicular stomatitis virus（VSV）. Methods Wish cells were transfected with pcDNA3.1-MxA（wild-type or L612K mutant）and control DNA respectively. After being infected with VSV for 48 hours, the amount of vial cells was determined by MTT. Subsequently, VSV mRNA level was detected by RT-PCR after wish cells were infected with VSV for 24 hours. Results The expression of wild-type MxA protein and L612K mutant was positively detected in wish cells. The amount of vial cells in L612K mutant group detected by MTT were found to be markedly low compared with that in wild-type and VSV mRNA level in L612K mutant group detected by RT-PCR which suggested markedly high compared with that in wild-type. Conclusion L612K mutant MxA showed the decrease of antiviral activity against VSV.