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Online Journal of Bioinformatics ©
Volume 11 (1): 162-176, 2010
Full-length DNA cloning, sequence analysis and transmembrane topology modelling of putative
Trypanosoma brucei rhodesiense oligosaccharyl transferase (TbOST I)
Waren N. Baticados1*(DVM, PhD), Noboru Inoue2 (DVM,PhD), Chihiro Sugimoto3 (DVM, PhD),
Hideyuki Nagasawa2 (DVM, PhD) and Abigail M. Baticados1 (DVM)
1College of Veterinary Medicine,University of the Philippines Los Baños, Laguna; 2National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 8555, Japan and 3 Laboratory of Infectious Diseases, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan.
Baticados WN, Inoue N, Sugimoto C, Nagasawa H, Baticados AM. Full-length DNA Cloning, Sequence Analysis and Transmembrane Topology Modelling of Putative Trypanosoma brucei rhodesiense Oligosaccharyl transferase (TbOST I), Onl J Bioinform, 11 (1): 162-176, 2010. The significant role played by glycans in altering a proteins antigenic properties and outcome of parasite evasion of the host defense mechanisms became the impetus for the full-length DNA cloning of Trypanosoma brucei rhodesiense oligosaccharyl transferase gene, STT3 subunit in particular. A gene with a coding sequence of 2430 bp and encoding a polypeptide of 809 amino acid residues was obtained. Preliminary sequence analysis revealed that homologues within the genome of other trypanosomatids and various origins exist. The conserve domain analysis showed that TbOST I belongs to OST STT3 subunit protein family and also possesses a WWDYG sequence motif ubiquitously found in STT3 homologues from various origins. Computational tool to predict topology was able to confirm that TbOST I is a putative membrane protein composed of 11 transmembrane regions with short N-terminal and long C-terminal domains located in the cytosol and lumen respectively.The paper reports an oligosaccharyl transferase STT3 subunit gene candidate from Trypanosoma brucei rhodesiense. This data also presented profound implications regarding the potential of TbOST I as putative drug target candidate against trypanosomosis. The TbOST I full-length nucleotide sequence has been deposited in a public database [GenBank: GU245936].
Key words: Trypanosoma brucei rhodesiense, oligosaccharyl transferase, N-glycosylation, STT3 subunit, Inverse PCR.