MOLECULAR DETECTION OF HEXOKINASE 1 GENE IN THE BLOODSTREAM FORM OF TRYPANOSOMA BRUCEI BRUCEI (FEDERE ISOLATE)
Ishaya Y. Longdet1* and Rotimi J. Ojo1,2
1Department of Biochemistry, University of Jos, Plateau State, Nigeria
2Department of Biochemistry, Bingham University, Karu, Nasarawa
email: islongdet@yahoo.com; longdeti@unijos.edu.ng
ABSTRACT
Hexokinase 1 (EC:2.7.1.1) which belongs to the Hexokinase 2 superfamily is the first key regulatory enzyme in the glycolytic pathway of Trypanosoma brucei brucei. Therefore the enzyme has great significance in metabolism of the parasite particularly for the purpose of generating ATP and nucleic acid precursors. There is increasing interest in the study of enzymes in parasites for various purposes. This research therefore attempted to detect the presence of the hexokinase 1 gene in Trypanosoma brucei brucei (Federe isolate) which is prevalent in Nigeria. The parasites were grown in rats, harvested and purified using diethyl amino ethyl (DEAE) cellulose chromatography. From the genomic DNA isolated the hexokinase 1 gene was amplified through Polymerase Chain Reaction. The amplicon was sequenced and studied using some Bioinformatics tools. The 1401bp gene sequence (Accession number MH 198230) obtained translated into 464 amino acid sequence. The primary nucleotide sequence of the gene compares well with Trypanosoma brucei TREU 927 and T. brucei gambienseDAL972.The predicted molecular mass of the polypeptide as calculated using ExPASy is 51,918.50Da (~52kDa) giving the hexameric protein a molecular weight of around 311.5kDa which differs from the mammalian hexokinase The Isoelectric point (pI) was predicted as 9.64indicating that atneutral pH, the enzyme has a netpositive charge. These revealed that the T.b.bruceihexokinase 1 is active in the bloodstream form of the parasite.