Recombinant Protein Production: A Case Study Research Report

Plasmodium spp. and Leishmania are the causing parasites to malaria and leishmaniasis, two widely spread diseases that are still responsible for millions of cases worldwide every year. The proteins in study, AGAP007752-PA (Anopheles gambiae) and ¿-tubulin (Plasmodium spp. and Leishmania infantum), are thought to be relevant in decreasing parasite transmission by affecting their life cycles. The aim is to use recombinant versions of those proteins to develop new control strategies to diminish disease incidence. In order to further the AGAP007752-PA research, the gene was amplified, cloned and sequenced. After, all three proteins were expressed in E. coli, and the recombinant proteins analysed by SDS-PAGE and Western Blot, again verifying their identities by sequencing. After the procedures¿ optimization, AGAP007752 gene amplification was achieved, although this protein was not successfully expressed. Contrarily, alfa-tubulins had been previously cloned, and the proteins¿ expression was now achieved. The research can be furthered, either through the production of the recombinant protein (AGAP007752-PA) or through purification of the expressed recombinant ¿-tubulin proteins.