Enhancement of Dengue-2 E Protein Expression by the Expression of the Precursor Membrane Protein (Prm) of the Dengue-3 Virus
Michelle D de Oliveira1, André S de Oliveira1, Nina R Dutra1, Rafael F O França2, Eduardo R Honda3, Fernando B Zanchi3, Clovis A Neves1 Cynthia C da Silva1, Benedito A L da Fonseca4 and Sérgio O De Paula1*
- *Corresponding Author:
- Sérgio Oliveira de Paula
Laboratory of Molecular Immunovirology
Federal University of Viçosa, Av. PH Rolfs
s/n., Viçosa, 36571-000, Minas Gerais, Brazil
E-mail: [email protected]
Received date: March 10, 2013; Accepted date: April 23, 2013; Published date: April 25, 2013
Citation: de Oliveira MD, de Oliveira AS, Dutra NR, França RFO, Honda ER, et al.(2013) Enhancement of Dengue-2 E Protein Expression by the Expression of the Precursor Membrane Protein (Prm) of the Dengue-3 Virus. J Vaccines Vaccin 4:182. doi: 10.4172/2157-7560.1000182
Copyright: © 2013 de Oliveira MD, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Several attempts to develop dengue recombinant subunit vaccines have failed due to insufficient levels of expression and incorrect folding of the E protein. In order to verify the importance of the precursor membrane protein to the level of E protein expression, we constructed two recombinant plasmids by cloning the full-length sequence of the prM gene from the dengue-2 and dengue-3 virus strains into a plasmid that expresses a truncated version of the E protein of the dengue-2 virus. Next, we evaluated these two constructs for their effect on the levels of E protein expression in vitro. Our results showed that both plasmids provided a correct expression of the E protein as E protein expression was detected in transfected Vero cells as demonstrated by indirect immunofluorescence and immunoblotting. Densitometry analysis of western blots of the cell extracts showed a 67.02% higher expression of E protein in the cells transfected with pCID2EtD3prM, indicating that the prM sequence of the dengue-3 virus may be more effective in assisting with the correct processing of the E protein. The results of this study may be used to increase the in vitro production of the E protein antigen for use in vaccines and for diagnostic purposes.