Expression of group c rotavirus VP6 protein in bacterial, baculoviral and Herpes virus based systems

ROTA, R.P.; ARGÜELLES, M.H.; LOZANO, M.E.; CASTELLO, A.A.; GLIKMANN, G.

Puerto Madryn

Congreso; XLVI Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2010

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

Rotavirus belongs to the Reoviridae family. The viral particle possesses a structural protein named VP6 that is of particular interest, since it forms part of the internal capsid, is not glycosylated and is the most inmunogenic of the structural proteins of the virion. In addition, based on the antigenic and genetic similarities of VP6 these viruses have been classified in groups A-G. Group C rotaviruses were detected in cases of diarrhea in humans and it has been demonstrated that infection and disease rates due to this agent are underestimated. It is necessary to emphasize that the lack of commercially available methods to detect the virus greatly contributes to this situation. In this context, the immunogenicity of VP6 protein and his preserved aminoacidic sequence turn it into a potentially excellent diagnostic tool. Specific primers were designed and the codifying region of human group C rotavirus VP6 gene was amplified by RT-PCR. The original material was clinical samples available in our laboratory. The amplified fragments were cloned in three different vectors according to the expression system to be used (bacterial, baculoviral and a herpes virus based system). The expression level of VP6 protein was evaluated by SDS-PAGE, Western blot and ELISA. This recombinant protein could be used in the development of diagnostic methods to detect this virus in diarrheal samples