Analysis of the protein expression level of the erythrocyte binding proteins in Plasmodium falciparum field isolates.

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Erythrocyte binding proteins are potent malaria vaccine candidates and the expression of these proteins in the field isolates has to be evaluated before vaccine development based upon these proteins. The objective of this project is to analyze mRNA and protein expression for the erythrocyte binding proteins, encoded by multigene family EBL, RBL, and Clag/RhopH1, which are involved in the erythrocyte invasion process. We collected P.falciparum field isolates in Thailand and cultured for short period and enriched mature schizonts. Using oligonucleotide targeting members of each multigene family, RT-PCR was performed to detect transcripts level. However, by preliminal analysis of the polymorphism of these genes, we found these gene family members are highly diversified and carefull design of the oligonucleotide is required to distinguish each member, but identify individual members. We generated a panel of antisera against each members and analyzed the expression levels of the each member. For Clag/RhopH1, the region showing diversified each other among members shows also polymorphic among isolates, indicated that it is required to generate antisera which distinguish each members, but react all type of the allele.