Expression studies of malarial antigen

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Date
2004
Authors
Ling, Rita Ting
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Publisher
Pusat Pengajian Sains Kesihatan, Universiti Sains Malaysia
Abstract
Serine repeat antigen (SERA) of Plasmodium falciparum is believed to be an excellent candidate for the development of a malaria vaccine. In this study, a 22-kDa synthetic DNA fragment (SE22) from the N-terminal domain of SERA which was previously constructed by assembly polymerase chain reaction (PCR) was subcloned from the cloning vector, pCR®2.1-TOPO® into an expression vector, pPRoEX™HTa for its expression in BCG. Prior to this study, the SE22 gene was constructed in favour of mycobacterium codon usage and cloned into pCR®2.1-TOPO® to produce pNMN009. In order to express the protein, the SE22 gene was extracted from pNMN009 and subcloned into pPROEX™HTa to produce pNMN019. The expression of the fusion protein was induced with isopropylthiogalactoside (IPTG). The fusion protein was isolated and used to immunize two New Zealand white rabbits to produce polyclonal antibody against the candidate antigen. This antibody will be used to detect the expression of the synthetic SE22 gene in Mycobacterium bovis bacille Calmette Guerin.
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Keywords
malaria
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