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目的初步确定汉坦病毒GM04-38株包膜糖蛋白上潜在的融合肽区域。方法采用基因定点突变技术将潜在融合肽区域的十个关键氨基酸突变成性质相左的氨基酸,转染Vero E6细胞后,间接免疫荧光(IFA)检测糖蛋白表达,采用Giemsa染色法观察细胞融合现象。结果在Vero E6细胞中成功表达出糖蛋白,IFA显示关键氨基酸突变前后细胞中均有荧光信号,呈胞浆分布,但细胞融合现象在突变后消失。结论潜在融合肽区域的十个氨基酸突变均对细胞融合产生明显的影响,提示该段区域很可能是病毒的融合肽。
Objective To determine the potential fusion peptide region of the envelope glycoprotein of Hantavirus GM04-38 strain. Methods Ten amino acids of potential fusion peptide were mutated into amino acids with opposite phase by gene mutagenesis. After transfection with Vero E6 cells, the expression of glycoprotein was detected by indirect immunofluorescence (IFA), and the cell fusion was observed by Giemsa staining . Results The glycoprotein was successfully expressed in Vero E6 cells. IFA showed the fluorescence signals in the cells before and after the key amino acid mutations, showing cytoplasmic distribution. However, the cell fusion phenomenon disappeared after mutation. Conclusion All ten amino acid mutations in the potential fusion peptide region have a significant effect on cell fusion, suggesting that this region is likely to be a viral fusion peptide.