Isolation, cloning and expression of the gene encoding lactate dehydrogenase enzyme from Theileria parva

Theileria parva is the causative agent of East Coast fever (ECF), a tick born disease, result in major economic losses in cattle. The high costs of drugs used to treat of ECF, development of resistance to some of these drugs , the absence of an effective vaccine against the parasite are significant problems in dealing with this illness. Thus, it is necessary that exploiting new resources to identify effective therapeutic targets to help develop new, effective, and cheap drugs, is imperative.Glycolysis is the main pathway of Theileria parva for energy production. The enzymes that play an important role in glycolysis may provide new targets for designing antitheilerials. Given its highest activity among glycolytic enzymes of the parasite, lactate dehydrogenase enzyme has been selected as the therapeutic target. Lactate dehydrogenase gene was cloned into pGEM-T vector with standard methods. TpLDH gene contains 2 introns, 3 exons and the gene is 1353 base pair in lenght. The introns were removed by site-directed mutagenesis then, exons fragments were ligated by PCR which generated a protein coding region 966 bp in length.Key Words: Theileria parva, Apicomplexan, Lactate Dehydrogenase, Antiheilerial, Chemotherapeutic Target