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Diseases of Aquatic Organisms

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DAO 61:33-39 (2004)  -  doi:10.3354/dao061033

Characterization of the 23S and 5S rRNA genes and 23S-5S intergenic spacer region (ITS-2) of Photobacterium damselae

Carlos R. Osorio1,*, Matthew D. Collins2, Jesús L. Romalde1, Alicia E. Toranzo1

1Departamento de Microbioloxía e Parasitoloxía e Instituto de Acuicultura,Universidade de Santiago de Compostela, 15782 Santiago de Compostela, Galicia, Spain
2School of Food Biosciences, University of Reading, PO Box 226, Whiteknights, Reading RG6 6AP, UK

ABSTRACT: The 23S ribosomal RNA (rRNA) gene has been sequenced in strains of the fish pathogens Photobacterium damselae subsp. damselae (ATCC 33539) and subsp. piscicida (ATCC 29690), showing that 3 nucleotide positions are clearly different between subspecies. In addition, the 5S rRNA gene plus the intergenic spacer region between the 23S and 5S rRNA genes (ITS-2) were amplified, cloned and sequenced for the 2 reference strains as well as the field isolates RG91 (subsp. damselae) and DI21 (subsp. piscicida). A 100% similarity was found for the consensus 5S rRNA gene sequence in the 2 subspecies, although some microheterogeneity was detected as inter-cistronic variability within the same chromosome. Sequence analysis of the spacer region between the 23S and 5S rRNA genes revealed 2 conserved and 3 variable nucleotide sequence blocks, and 4 different modular organizations were found. The ITS-2 spacer region exhibited both inter-subspecies and inter-cistronic polymorphism, with a mosaic-like structure. The EMBL accession numbers for the 23S, 5S and ITS-2 sequences are: P. damselae subsp. piscicida 5S gene (AJ274379), P. damselae subsp. damselae 23S gene (Y18520), subsp. piscicida 23S gene (Y17901), P. damselae subsp. piscicida ITS-2 (AJ250695, AJ250696), P. damselae subsp. damselae ITS-2 (AJ250697, AJ250698).


KEY WORDS: 23S · 5S · ITS-2 · Photobacterium damselae · Intergenic spacers


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