Ling, Rita Ting
(2004)
Expression studies of malarial
antigen.
Expression studies of malarial antigen.
(Submitted)
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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