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Cloning and characterization of ribosomal RNA genes from Opisthorchis viverrini

Published online by Cambridge University Press:  06 April 2009

S. Korbsrisate ,
S. Mongkolsuk ,
J. R. Haynes ,
Q. Wong  and
S. Sirisinha
S. Korbsrisate
Affiliation:
Departments of Microbiology, Faculty of Science, Mahidol University, Bangkok, Thailand
S. Mongkolsuk
Affiliation:
Biotechnology, Faculty of Science, Mahidol University, Bangkok, Thailand
J. R. Haynes
Affiliation:
Division of Molecular Genetics, Connaught Centre for Biotechnology Research, Willowdale, Ontario, Canada
Q. Wong
Affiliation:
Division of Molecular Genetics, Connaught Centre for Biotechnology Research, Willowdale, Ontario, Canada
S. Sirisinha
Affiliation:
Departments of Microbiology, Faculty of Science, Mahidol University, Bangkok, Thailand
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Summary

The ribosomal DNA (rDNA) unit of the liver fluke Opisthorchis viverrini has been cloned and characterized. The results demonstrated that the total length of this unit is approximately 13 kb, containing 4·2 kb of large subunit (LSU) rDNA, 2·0 kb of small subunit (SSU) rDNA, 1·0 kb of transcribed spacer DNA and 5·8 kb of non-transcribed + external transcribed spacer DNA. Examination of the non-transcribed spacer region between different rDNA units showed variation in the restriction sites rather than in the length. Judging from hybridization of the rDNA plasmid to the restriction endonuclease digest of genomic DNA, rDNA units represent 6·1% of the total genomic DNA. At the RNA level, the LSU rRNA of O. viverrini and Fasciola gigantica contained a hidden break. The molecule consisted of two fragments co-migrating with a SSU rRNA, when electrophoresis was carried out under denaturing conditions. Ribosomal RNA sequence comparison has been previously used to determine phylogenetic classification of parasitic organisms. The sequence of 381 nucleotides at the 5′ terminus of the LSU rRNA gene was determined and compared with those from species previously reported by other investigators. Phylogenetic classification of O. viverrini, as determined by rRNA gene sequence comparison, is comparable with the conventional taxonomic classification.

Keywords

Opisthorchis viverrinirRNArDNAhidden breakcopy numberphylogeny
Type
Research Article
Information
Parasitology , Volume 104 , Issue 2 , April 1992 , pp. 323 - 329
Copyright
Copyright © Cambridge University Press 1992

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