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Shi, Ya Ping and Das, P. and Holloway, B. and Udhayakumar, V. and Tongren, J.E. and Candal, F. and Biswas, S. and Ahmad, S.R. and Hasnain, S.E. and Lal, A.A. (2000) Development, expression, and murine testing of a multistage Plasmodium falciparum malaria vaccine candidate. Vaccine, 18 (25). pp. 2902-2914. ISSN 0264-410X

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A synthetic gene encoding twelve B cell epitopes, six T-cell proliferative epitopes, and three cytotoxic T lymphocyte (CTL) epitopes from nine stage-specific antigens, representing the sporozoite, liver stage, asexual blood-stage, and sexual-stage antigens of Plasmodium falciparum, was constructed by assembling overlapping oligonucleotides followed by PCR extension and annealing. A three-step PCR protocol using twelve long oligonucleotides was employed to generate a 1053 base-pair synthetic gene, the identity of which was confirmed by sequencing. This synthetic gene, named CDC/NII MAL VAC-1, was cloned, and the recombinant protein was expressed in the Baculovirus Expression Vector System (BEVS). The selection of malarial epitopes for inclusion in this vaccine construct was based on immunoepidemiological studies in malaria endemic area, in vitro, and in vivo protection studies in model systems. The 41 kDa BEVS-expressed recombinant protein reacted with mouse antibodies specific for individual B cell epitopes in the vaccine construct and with sera from clinically immune Kenyan adults. An immunization study in three strains of mice that differ at the H-2 locus demonstrated that the BEVS-expressed recombinant protein is immunogenic; the candidate vaccine antigen induced high titer antibodies, and lymphocyte proliferative and IFN-γ responses. These results demonstrate that individual B and T cell epitopes can be assembled to create synthetic genes that encode proteins capable of eliciting specific antibody and T cell responses.

Item Type: Article
Depositing User: Dr P Divakar
Date Deposited: 28 May 2015 07:35
Last Modified: 28 May 2015 07:35
URI: http://cdfd.sciencecentral.in/id/eprint/100

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