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OJVR

Online Journal of Veterinary Research©

 Volume 14 (1):56-65, 2010. Redacted 2017.


Molecular cloning and sequencing of Trypanosoma brucei rhodesiense putative oligosaccharyl transferase

 

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.


ABSTRACT

 

Baticados WN, Inoue N, Sugimoto C, Nagasawa H, Baticados AM Molecular Cloning and sequencing of Trypanosoma brucei rhodesiense Putative Oligosaccharyl transferase, Onl J Vet Res., 14 (1):56-65, 2010.  Genomic clone of putative Trypanosoma brucei rhodesiense oligosaccharyl transferase (TbrOST I) with ~2000 bp DNA size was obtained by PCR amplification. The size of the amplicon is akin to other kinetoplastid homologues.  Sequencing revealed a partial nucleotide sequence of 2048 bp encoding a predicted protein sequence of 682 amino acid residues. TbrOST I has a very high nucleotide (Ns) and amino acid (As) sequence identity to putative oligosaccharyl transferase subunit in the genomes of Trypanosoma brucei gambiense (Ns:98%; As:99%) and T. brucei (Ns:94-98%; As:93-99%).  However, only a moderately high nucleotide identity and low amino acid sequence similarity to putative oligosaccharyl transferase subunit was observed in the genomes of Trypanosoma cruzi (Ns:74%; As:61%), Leishmania infantum  (Ns:70-80%; As:46-54%),  L. braziliensis (Ns:68-78%; As:46-55%) and L. major  (Ns:69-76%; As:45-54%). Moreover, TbrOST I also exhibited moderately high sequence identity (71-77%) to the genome from other origins. Based on the size of amplicon and high percent identity to reported or putative oligosaccharyl transferase subunit in the genome of related species and a variety of organisms, the results strongly indicate that TbrOST  I is a potential trypanosome oligosaccharyl transferase gene. The sequence data of TbrOST I has been deposited in the GenBank with accession number GU475125.

 

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


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