Expression of NSP 3AB Gene of Encephalomyocarditis Virus(EMCV)in E.coli and Development of Monoclonal Antibodies Against 3AB Protein
- VernacularTitle:猪脑心肌炎病毒非结构蛋白3AB的原核表达及其单克隆抗体的研制
- Author:
Fang SHEN
;
Ping JIANG
;
Yu-Feng LI
;
Jun-Xing LI
;
Hai-Yan WANG
;
- Publication Type:Journal Article
- Keywords:
Encephalomyocarditis virus(EMCV) Non-structural protein Prokaryotic expression Monoclonal antibody
- From:
China Biotechnology
2006;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To express the EMCV 3AB gene by prokaryotic expression systerm,and prepare monoclonal antibodies against it. Method: NSP 3AB gene of Encephalomyocarditis virus (EMCV) was amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a recombinant protein 3AB with high antigenicity was expressed in E.coli. Balb / c mice were immunized by purified recombinant 3AB protein of inclusion-body, and the splenocytes of the immunized mice were fused with murine myeloma cells to produce hybridoma cell line. Results: After subcloning by 3 times, one strain of hybridoma cell line steadily secreting antibodies of 3AB protein was obtained, named 2D12. The McAb belongs to IgG1/?. The McAb and was confirmed by indirect immunofluorescent assay (IFA) and Western blot. Conclusion: These results can provide a potential value for structural and functional studies of EMCV-3AB and early diagnosis of Encephalomyocarditis virus infection.
