Demonstration of IS711 transposition in Brucella ovis and Brucella pinnipedialis.
BACKGROUND
The Brucella genome contains an insertion sequence (IS) element called IS711 or IS6501, which is specific to the genus. The copy number of IS711 varies in the genome of the different Brucella species, ranging from 7 in B. abortus, B. melitensis and B. suis to more than 30 in B. ovis and in Brucella strains isolated from marine mammals. At present, there is no experimental evidence of transposition of IS711, but the occurrence of this element with a high copy number in some species, and the isolation of Brucella strains with "ectopic" copies of IS711 suggested that this IS could still transpose.


RESULTS
In this study we obtained evidence of transposition of IS711 from the B. ovis and B. pinnipedialis chromosomes by using the "transposon trap" plasmid pGBG1. This plasmid expresses resistance to tetracycline only if the repressor gene that it contains is inactivated. The strains B. melitensis 16 M, B. abortus RB51, B. ovis BOC22 (field strain) and B. pinnipedialis B2/94, all containing the plasmid pGBG1, were grown in culture media with tetracycline until the appearance of tetracycline resistant mutants (TcR). TcR mutants due to IS711 transposition were only detected in B. ovis and B. pinnipedialis strains.


CONCLUSIONS
Four different copies of IS711 were found to transpose to the same target sequence in the plasmid pGBG1. This demonstrated that IS711 are active in vivo, specially in Brucella species with a high number of IS711 copies as B. ovis and B. pinnipedialis.
Ocampo-Sosa
Alain A
AA
Departamento de Biología Molecular, Universidad de Cantabria, Instituto de Biomedicina y Biotecnología de Cantabria, IBBTEC, CSIC-Universidad de Cantabria-IDICAN, Santander, Spain. alainocampo@hotmail.com
García-Lobo
Juan M
JM
D015139:Blotting, Southern; D002002:Brucella / Q000235:genetics*; D042061:Brucella ovis / Q000235:genetics*; D004251:DNA Transposable Elements / Q000235:genetics*; D016680:Genome, Bacterial; D016254:Mutagenesis, Insertional; D009154:Mutation; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D004251:DNA Transposable Elements