Biotechnology Journal International

Aim: This work focused on the sequence homology studies of the enzyme, phospholipase A2  (PLA₂), in Trypanosoma brucei  obtained from the blood of bull  in Federe, Plateau State, Nigeria, West Africa

Place and Duration of Study: Department of Biochemistry, University of Jos, Nigeria; Department of Biochemistry, Ahmadu Bello University, Zaria, Nigeria, Department of Biotechnology, NVRI, Vom, Nigeria; between June 2009 and September 2011.

Methodology: T. brucei grown in rats were harvested and separated using diethyl amino ethyl (DEAE) cellulose chromatography. From the parasites’ genomic DNA the PLA₂-like gene was amplified using consensus primers.  The amplicon was cloned unto pMal-2cE vector and confirmed using direct PCR and restriction enzyme analyses. The PLA₂ gene and translated protein sequences were studied using National Center for Biotechnology Information (NCBI) Conserved Domain Search Tool and Conserved Domain Architectural Retrieval Tool

Results: Analyses of the 1344bp gene sequence using bioinformatics tools showed that it is very closely related to PLA₂ sequences of T. brucei (TREU 927) and T. b. gambiense. Motifs that are unique to PLA₂ (FSHGL) and lipases (GHSFG) were found to be present in the query sequence. The domains present in the studied sequence agreed closely with those of the human platelet activating factor acetyl hydrolase (PAF-AH). There was also a good sequence resemblance with PLA₂s from T. cruzi, Metarhizium amisop, Metarphizium acridu and PAF-AH in terms of architecture.

Conclusion: The PLA₂-like gene isolated from the blood stream form of Trypanosoma brucei and studied was found to posses the domains and motifs unique to PLA₂s and lipases and so homology was established among the proteins.