title: Mapping of IS6110 flanking regions in clinical isolates of Mycobacterium tuberculosis demonstrates genome plasticity.
abstract: Southern hybridization was used in combination with IS6110 insertion-locus-specific probes in a comparative study to determine the structure of chromosomal domains flanking IS6110 elements in clinical isolates of Mycobacterium tuberculosis. The resulting restriction fragment length polymorphism (RFLP) data demonstrated three mutational mechanisms responsible for the polymorphisms observed: IS6110 insertion, chromosomal mutation and deletion. The frequency of IS6110 insertion within many of the chromosomal regions demonstrates that preferential integration regions are common in M. tuberculosis. Mapping the IS6110 insertion positions and chromosomal deletions in relation to the M. tuberculosis H37Rv and M. bovis BCG genome sequences reveals numerous disruptions of predicted open reading frames (ORFs). A phylogenetic tree, based on the mutational data, showed a number of independently evolving lineages of M. tuberculosis, while analysis of the mutational events occurring at each branch point suggests both divergent and convergent evolution. A significant positive correlation was demonstrated between the mutation rate and the frequency of occurrence of different isolates in families of strains, suggesting that evolution may impact on strain 'fitness' or that strain proliferation may increase the chance of mutation. We conclude that the genome of clinical isolates of M. tuberculosis continues to evolve.
authors.affiliation: MRC Centre for Molecular and Cellular Biology, Department of Medical Biochemistry, University of Stellenbosch, PO Box 19063, Tygerberg, 7505, South Africa.
authors.forename: R M
authors.initials: RM
authors.lastname: Warren
authors.forename: S L
authors.initials: SL
authors.lastname: Sampson
authors.forename: M
authors.initials: M
authors.lastname: Richardson
authors.forename: G D
authors.initials: GD
authors.lastname: Van Der Spuy
authors.forename: C J
authors.initials: CJ
authors.lastname: Lombard
authors.forename: T C
authors.initials: TC
authors.lastname: Victor
authors.forename: P D
authors.initials: PD
authors.lastname: van Helden
mesh_terms: D005075:Biological Evolution; D015139:Blotting, Southern; D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial; D004251:DNA Transposable Elements; D016680:Genome, Bacterial; D006801:Humans; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000235:genetics*; D010802:Phylogeny; D012150:Polymorphism, Restriction Fragment Length
chemical_list: D004251:DNA Transposable Elements