Transfecting Plasmodium berghei with the Plasmodium falciparum thiazole kinase gene
Abstract
As current antimalarial drugs become progressively ineffective due to parasite resistance
there is need to develop and pursue new therapeutic strategies. Parasite-specific metabolic
pathways promise to make an ideal source of novel drug targets and provide unique
opportunities for chemotherapy. Vitamin B1 biosynthesis pathway in Plasmodium falciparum
is one such pathway. The overall aim of this study was to transfect Plasmodium berghei to
express the Plasmodium falciparum thiazole kinase (Pfthk) gene. Following successful DNA
isolation and peR, a 909 base pair gene fragment was isolated from Plasmodium falciparum
genomic DNA. Upon expression of the gene in bacterial systems, the protein was found to be
soluble with a molecular weight of 34.67 kDa. The thk was successfully cloned into a P.
berghei expression vector, pExpress-l thus generating a stable transfection construct.
Parasites (2.5xl09) were obtained from Balb/c mice (with a 5% parasitaemia of schizont
stage) and upon overnight in vitro culture, the schizont stage grew to 5.0x I09 parasites. For
transfection of P. berghei blood-stage parasites, electroporation settings of I Kv, 25~lF and
200 Ohms were used. Time constants of 0.7 rns and 0.8 rns were attained for two samples,
showing a successful electroporation. Analysis of data showed that PFTHK protein is soluble
and the amino acid signal peptide analysis revealed that the protein was non-secretory and
without a cleavage site. These are characteristics of an attractive drug target. Further studies
in the development of the protein as a drug target are recommended.
Citation
Master ofSeienee in Applied ParasitologySponsorhip
University of NairobiPublisher
University of Nairobi School of Biological Sciences