Expression and purification of full-length recombinant Plasmodium falciparum PfMC-2TM Maurer’s cleft protein

Le résumé fourni par la source

Malaria caused by Plasmodium falciparum remains the most virulent form of malaria, resulting in 216 million cases and 445,000 deaths globally. Invasion of red blood cells by P. falciparum leads to the formation of membranous structures known as Maurer’s clefts (MC). Virulence markers of P. falciparum such as PfEMP1 are transported across the MC to the surface of the infected red blood cell. Insight into the formation and function of the MC will be important for the discovery of new vaccine and drug candidates. The PfMC-2TM is encoded by a multi-gene family of 13 members. PfMC- 2TM is a protein localized to the MC. We induced expression of PfMC-2TM encoded by 1 family member [PF3D7_0114100 (PFA0680c)] in BL21 DE3 strain of Escherichia coli following transformation with recombinant pET-28a plasmid containing a chemically synthesized gene. The purpose of this study was to determine immunogenic properties of the resulting recombinant protein using western blot analysis. The recombinant plasmid was isolated and analyzed in 1% agarose gel and an approximately 5kb band was identified. Pilot expression of transformants showed expression of recombinant PfMC-2TM by western blot. Recombinant PfMC-2TM protein will be expressed and purified for antibody production to allow subsequent domain analysis and characterization.

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