Construction and verification of recombinant vesicular stomatitis virus expressing tick-borne encephalitis virus prM-E protein
10.13200/j.cnki.cjb.004668
- VernacularTitle:表达森林脑炎病毒prM-E蛋白重组水疱性口炎病毒的构建及鉴定
- Author:
Yuying NING
1
Author Information
1. Yan’an Medical College of Yan’an University, Yan’an 716000, Shaanxi Province, China
- Publication Type:Journal Article
- Keywords:
Vesicular stomatitis virus(VSV);
Tick-borne encephalitis virus(TBEV);
prM-E protein;
Reverse genetics
- From:
Chinese Journal of Biologicals
2026;39(03):264-269+276
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo construct a recombinant virus expressing tick-borne encephalitis virus(TBEV) prM-E protein using vesicular stomatitis virus(VSV) as a vector, and to identify it, so as to provide a basis for the research of TBEV vaccines based on VSV vector.MethodsThe prM-E gene of the TBEV Senzhang strain was inserted between the G and L genes of the VSV vector. The recombinant virus was rescued by co-transfecting BHK-21 cells infected by poxvirus containing T7 RNA polymerase with helper plasmids expressing VSV N, P, L, and G proteins. The supernatant was collected, and the recombinant virus rVSV-TBEVprM-E stably expressing prM-E was obtained after multiple passages. Western blot, immunofluorescence assay(IFA) and RT-PCR were used to identify the expression of prM-E protein and gene of rVSV-TBEVprM-E in cells. The viral titers of rVSV-TBEVprM-E at different time points were determined by plaque assay and the growth curve was plotted.ResultsTBEV prM-E gene was successfully inserted into the genome of recombinant virus rVSV-TBEVprM-E, and the expression of prM-E protein in BHK-21 cells was detectable. After serial passages, rVSV-TBEVprM-E achieved a viral titer of 6. 75 × 10~5 PFU/mL.ConclusionA recombinant virus rVSV-TBEVprM-E expressing prM-E protein was successfully constructed, which lays a solid experimental foundation for the related research of TBEV.
