汉坦病毒GM04-38株包膜糖蛋白上融合肽的初步确定

Defining fusion peptide on glycoprotein of the Hantavirus GM04-38

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 putative fusion region on glycoprotein of the Hantavirus GM04-38. Methods Site-directed mutagenesis was used to construct 10 GP gene mutants, which were then expressed in Vero E6 cells. IFA was performed to determine expression of GPs genes, and Giemsa staining was used to observe cell fusion activities. Results IFA showed that the GPs genes were successfully expressed, and the fluorescent signal was robust and concentrated in the perinuclear region of the transfected cells. But cell fusion activities were not found after mutation. Conclusion Mutation of 10 amino acids can obviously influence cell fusion activities, suggesting that it is possible that fusion peptide is in this region.