Expression and
Purification of Recombinant Malarial Antigen Plasmodium falciparum Lactate Dehydrogenase
(PfLDH) in E.Coli Expression System for Diagnostic Application
Krithiga K., Sivarathnakumar S., Praveen kumar R. and Vinoth Arul Raj J.
Res. J. Biotech.; Vol. 20(7); 150-155;
doi: https://doi.org/10.25303/207rjbt1500155; (2025)
Abstract
Malaria is still a fatal endemic disease caused by plasmodium parasites, most commonly
transmitted by human bites transmitted by female Anopheles mosquitoes. It remains
one of the biggest global threats to human health, especially in subtropical areas.
The main stage of the parasite (Plasmodium falciparum) is responsible for most of
those severe and lethal cases of malaria. The stage of the parasite's life cycle
when it infects and replicates the phase occurring inside the host organism’s red
blood cells, is typically a human, called the erythrocytic stage. Plasmodium falciparum
Lactate Dehydrogenase (PfLDH) is a glycolytic enzyme that catalyses the conversion
of pyruvate and lactate into energy for the body's metabolism. It is used in antimalarial
medications and diagnostic tests for finding malaria because the presence of PfLDH
in blood samples suggests the presence of the parasite. The focus of this study
is to make recombinant PfLDH protein in Escherichia coli BL21(DE3) strain.
The PfLDH gene was introduced into the expression vector pET27b(+)to construct recombinant
plasmid, which was then transformed into E. Coli BL21(DE3). Next, recombinant DNA
containing host cell was induced by IPTG at different concentrations, time intervals
and temperature that can be grown in LB broth containing marker gene. Recombinant
protein PfLDH was purified by IMAC, was analysed and was confirmed by SDS-PAGE and
Western blot. It can be utilized as a biomarker for malarial Rapid Diagnostic Tests
(RDTs).
