汉坦病毒包膜糖蛋白糖基化位点突变体的构建

Construction of N-linked glycosylation site mutants of envelope glycoprotein genes of hantavirus

目的构建汉坦病毒糖基化位点的突变体。方法利用基因定点突变的方法构建了5个糖蛋白突变体,即将G1、G2上的天冬酰胺置换为丙氨酸,根据被替换的位置,突变体分别命名为N134A、N235A、N347A、N399A、N928A。结果构建的5个N-联糖基化位点的突变体,经测序图谱显示原序列中的天冬酰胺(N)均被置换为丙氨酸(A)。结论成功构建了5个糖基化位点的突变体,为进一步研究N-联糖基化的缺失对细胞融合的影响奠定了基础。

Objective To construct N - linked glycosylation site mutants of hantavirus. Methods Site - directed mutagenesis was used to construct five glycoprotein gene mutants which were designed as N134A, N235A, N347A, N399A and N928A according to the substitution sites of asparagine with alanine on G1 and G2. Results Five N - linked glycosylation site mutants were constructed and their sequencing showed the asparagine （N） residues at the N -linked glycosylation sites on G1 and G2 were replaced by alanine （A）. Conclusion N -linked glycosylation site mutants were successfully constructed and laid the foundation for study on the roles of N -linked glycosylation of HTNV glycoproteins in cell fusion.