Construction and verification of recombinant chimeric vesicular stomatitis virus expressing rabies virus G protein
10.13200/j.cnki.cjb.000079
- VernacularTitle:表达狂犬病病毒G蛋白嵌合型重组水疱性口炎病毒的构建及鉴定
- Author:
ZHAO Tingting
- Publication Type:Journal Article
- Keywords:
Vesicular stomatitis virus(VSV);
Rabies virus(RV);
Reverse genetics;
Chimeric virus
- From:
Chinese Journal of Biologicals
2023;36(12):1450-1454
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct recombinant chimeric vesicular stomatitis virus(VSV)expressing G protein of rabies virus(RV)using VSV as vector.MethodsTo rescue the recombinant virus,G gene of VSV antigenome was replaced with G gene of RV vaccine CTN-1 strain,and co-transfected into 293T cells with helper plasmids coding T7 RNA polymerase and proteins N,P and L of VSV. The expression of RV G gene and G protein was detected by RT-PCR,immunofluorescence assay and Western blot. The recombinant virus was subcultured in Vero cells,the virus titer of different generations was detected and the virus growth curve was drawn.ResultsThe recombinant virus VSV-RVG was successfully rescued. RTPCR results demonstrated that the RV G gene was successfully inserted into the genome of the recombinant virus,and the expression of RVG protein was detected by immunofluorescence assay and Western blot. The recombinant virus was continuously passaged for 5 generations,and the virus titer was stable within 7. 5 ～ 8. 5 lgTCID50/mL.ConclusionThe recombinant chimeric VSV expressing RV G protein was successfully constructed with good genetic stability,which lays a foundation of the construction of reverse genetics technology platform based on VSV vector.
