摘要
目的研究K83A变异对MxA蛋白抑制VSV复制活性的影响。方法将野生型和K83A变异MxA蛋白载体pcDNA3.1-MxA(WT和K83A)分别瞬时转染Wish细胞,24h后VSV感染细胞,感染48h后MTT检测各组细胞增殖;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VSVmRNA水平;Western blot检测各组MxA蛋白表达水平。结果野生型和K83A变异MxA蛋白均在Wish细胞有较好表达;MTT检测结果提示K83A组细胞增殖数明显低于WT组(P<0.01),RT-PCR结果显示K83A组VSVmRNA水平明显高于WT组(P<0.01),但与对照组比较差异无统计学意义(P>0.05)。结论K83A变异MxA蛋白使MxA蛋白失去了抑制VSV复制的活性。
Objective To investigate the antiviral activity of K83A mutant MxA protein against vesicular stomatitis virus. Methods The wish ceils in MxA groups were transfected with pcDNA3.1-MxA （ wild-type or K83A mutant） and control DNA respectively. After infected with VSV for 48h, the amount of vial cells was determined by MTT. Subsequently, VSV mRNA level was detected by RT-PCR after wish cells was transfected and infected with VSV for 24h. Results The expression of wildtype MxA protein and K83A mutant was positive in wish cells. The amount of vial cell in K83A mutant group detected by MTT showed markedly low compared with that in wild-type and the level of VSV mRNA in K83A mutant group detected by RT-PCR suggested markedly high compared with that in wild -type respectively. Conclusion K83A mutant MxA showed the deficiency of antiviral activity against VSV.
出处
《四川医学》
CAS
2009年第11期1694-1696,共3页
Sichuan Medical Journal
