Isolation and cloning of Plasmodium vivax lactate dehydrogenase gene by polymerase chain reaction

ABSTRACT MASTER THESIS ISOLATION AND CLONING OF Plasmodium vivax LACTATE DEHYDROGENASE GENE BY POLYMERASE CHAIN REACTION Ekrem AKBULUT Fırat University Graduate School of Natural and Applied Sciences Department of Biology 2003, Page : 50 Emergence of drug resistance to the existing antimalarials emphasizes the need for the development of new drugs. To this aim, in this thesis, a lactate dehydrogenase gene was cloned and sequenced from genomic DNA of Plasmodium vivax Belem strain (PvLDH). This is the second human malarial lactate dehydrogenase cloned to date. DNA sequence showed that like Plasmodium falciparum LDH (P/LDH), PvLDH contains a 951 bp open reading frame, coding for a 3 16 aminoacid protein. Aminoacid sequence alignment of PvLDH to P/LDH and to some other known LDHs indicated that PvLDH has %90,1 residue identities with P/LDH and 90,8%, 90,5%, 26%o, 25%), P. berghei, P. yoelii, insan-M4, insan-H4 LDHs respectively. It was also shown that PvLDH has two single aminoacid deletions, which are Glutamat-48 and Glisin-217. Most remarkable is a five residue insertion in the active site loop region. The high resolution X- ray structure previously obtained from P/LDH showed that this site might be an attractive target for inhibitors. Keywords : Malaria, Plasmodium vivax, Lactate Dehydrogenase, Polymerase Chain Reaction. IX