Molecular Cloning of Plasmodium Shikimate Kinase

Malaria is a disease that has a significant global burden and is both economically and clinically obstructive. New attempts to target the causative agent, the Plasmodium parasite, are focusing on the shikimate biosynthetic pathway due to both its role in producing key metabolites for the organism as well as its absence in animals. Our work focuses on the molecular cloning of the shikimate kinase gene, that codes for a key enzyme in this metabolic pathway, into a pEX-N-GST expression vector. This was carried out using P. falciparum and P. knowlesi shikimate kinase genes that were PCR amplified, restriction enzyme digested, ligated, and transformed into competent E. coli cells. It was found that the desired gene construct was not obtained potentially due to choice of expression vector, restriction enzymes, or ligation conditions. More work can be done to elucidate a successful cloning protocol and investigate other components of the shikimate biosynthetic pathway.