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Identification of novel DNA fragments and partial sequence of a genomic island specific of Brucella pinnipedialis

Maquart, Marianne, Fardini, Yann, Zygmunt, Michel S., Cloeckaert, Axel
Veterinary microbiology 2008 v.132 no.1-2 pp. 181-189
seals, whales, brucellosis, Brucella, molecular epidemiology, pathogen identification, microbial genetics, genome, genomics, molecular genetics, polymerase chain reaction, restriction mapping, sequence analysis, nucleotide sequences, DNA fragmentation, phylogeny
Since the 1990s, Brucella strains have been isolated from a wide variety of marine mammals and were recently recognized as two different species, i.e. Brucella pinnipedialis for pinniped isolates and Brucella ceti for cetacean isolates. The aim of this study was to identify specific DNA fragments of marine mammal Brucella strains using a previously described infrequent restriction site-PCR (IRS-PCR) method but with three new couples of restriction enzymes applied on a larger panel of marine mammal Brucella isolates (n =74) and one human isolate from New Zealand likely from marine mammal origin. This study revealed five DNA fragments specific of Brucella strains isolated from marine mammals. Among them two new DNA fragments were specific of B. pinnipedialis but were not detected in hooded seal isolates. DNA fragment I identified in the previous IRS-PCR study and fragment VI of this study were located on a cloned and sequenced 6kb SacI fragment. Its nucleotide sequence revealed that it is likely part of a putative genomic island. Sequence analysis showed that it carries four ORFs coding for putative metabolic functions. Although hooded seal isolates are classified within B. pinnipedialis it was shown in this study that they do not carry this genomic island and this raises the question about their evolutionary history within B. pinnipedialis.