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OJVRTM

 Online Journal of Veterinary Research © 

Volume 14 (2): 163-175, 2010

Molecular Characterization of Trypanosoma brucei rhodesiense Putative Oligosaccharyl Transferase STT3 subunit (TbOST I)

 

Waren N. Baticados1*(DVM, PhD), Noboru Inoue2 (DVM, PhD), Chihiro Sugimoto3 (DVM, PhD), Hideyuki  Nagasawa2 (DVM, PhD) and Abigail M. Baticados1 (DVM)

 

1Department of Veterinary Paraclinical Sciences, College of Veterinary Medicine, University of the Philippines Los Baños, Laguna, Philippines 4031., 2National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine,  Obihiro, Hokkaido 8555, Japan and 3Research Center for Zoonosis Control, Hokkaido University N18-W9, Kita-ku, Sapporo, Hokkaido 060-0818, Japan

ABSTRACT

 

Baticados WN, Inoue N, Sugimoto C, Nagasawa H, Baticados AM Molecular Characterization of Trypanosoma brucei rhodesiense Putative Oligosaccharyl Transferase STT3 subunit (TbOST I). Online J Vet Res 14 (2): 163-175, 2010.  Oligosaccharyl transferase functions as a catalyst for a complex carbohydrate-protein modification reaction, N-linked glycosylation, leading to the production of glycoproteins that contribute to its overall surface architecture.  The molecular cloning and sequencing of putative Trypanosoma brucei rhodesiense oligosaccharyl transferase candidate was previously reported. Subsequently, described in this paper is the molecular characterization of a novel Trypanosoma brucei rhodesiense putative oligosaccharyl transferase STT3 subunit (TbOST I). A single-copy gene of TbOST I was established after Southern blot analysis.  Northern hybridization revealed two (3.5 kb and 4.5 kb) mRNA transcripts in TbOST I. Immunoblot analysis exhibited that antibodies specific for the synthetic TbOST I oligopeptides reacted to ~50-55 kDa protein band.  Furthermore, immunoflorescense test indicates a cytoplasmic and flagellar localization of TbOST I in Trypanosoma brucei rhodesiense bloodstream forms.

 

 Key words:Trypanosoma brucei rhodesiense, oligosaccharyl transferase, N-glycosylation, STT3 subunit        


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