Construction of severe fever with thrombocytopenia syndrome virus recombinant pseudoviruses and effect of Gc glycosylation on viral infectivity
10.3760/cma.j.cn112866-20230712-00003
- VernacularTitle:SFTSV重组假病毒的构建及Gc糖基化对病毒感染力的影响
- Author:
Xiaowen CHONG
1
;
Zequn WANG
;
Mengting CHEN
;
Mengyu DU
;
Xiaoying XU
;
Youxiang MA
;
Hongling WEN
Author Information
1. 山东大学齐鲁医学院公共卫生学院微生物检验学系 山东省"十四五"高等学校新发突发传染病防控与生物安全重点实验室，济南 250012
- Keywords:
Severe fever with thrombocytopenia syndrome virus;
Gc;
Pseudovirus;
Glycosylation site
- From:
Chinese Journal of Experimental and Clinical Virology
2023;37(6):583-591
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the relationship between severe fever with thrombocytopenia syndrome virus (SFTSV) Gc and its N-glycosylation site and viral infectivity, a recombinant pseudovirus containing SFTSV Gc glycosylation site mutant was constructed.Methods:The eukaryotic expression vectors pcDNA3.1(+ )-GC, PCDNA3.1(+ )-GC(N291Q), PCDNA3.1(+ )-GC(N352Q) and PCDNA3.1(+ )-GC (N374Q) were constructed by site-directed mutagenesis and homologous recombination. After their successful expression in 293T cells, we infected VSVΔG-Fluc*G pseudovirus, constructed four recombinant pseudoviruses and tested their effects on the cell force of infection.Results:Double digestion identification and sequence determination confirmed the successful construction of eukaryotic expression vectors pcDNA3.1 (+ )-Gc, pcDNA3.1 (+ )-Gc(N291Q), pcDNA3.1 (+ )-Gc(N352Q) and pcDNA3.1 (+ )-Gc(N374Q). Indirect immunofluorescence and Western Blotting result indicated the successful expression of all the four recombinant plasmids. SFTSV Gc recombinant pseudoviruses are specific for infecting Vero cells. Pseudovirus infection capacity was decreased significantly after the glycosylation site mutation, and the mutant strain with the glycosylation site at position 352 had the lowest level of infectivity ( P<0.001, P=0.001). Conclusions:The glycosylation site of SFTSV Gc may be associated with the infectious effect of the viral infection, and the amino acid mutation at position 352 has the greatest effect on the viral infectivity.
