title: A novel polymorphic genetic locus in members of the Mycobacterium tuberculosis complex.
abstract: It has previously been shown that the PAN promoter from Mycobacterium paratuberculosis can be used as a DNA probe to identify an RFLP between wild-type Mycobacterium bovis and the vaccine strain Mycobacterium bovis BCG. To investigate the genetic basis of this phenomenon, DNA fragments from a New Zealand M. bovis cattle strain and M. bovis BCG Pasteur, containing the PAN-binding region, were isolated from gene libraries, sequenced and characterized. Sequence analysis and comparison with database sequences showed that the PAN region in M. bovis, M. bovis BCG and Mycobacterium tuberculosis is identical and shares 70% similarity to the PAN sequence from M. paratuberculosis. The Shine-Dalgarno sequence and the -10 and -35 promoter regions are conserved between the different species. Analysis of the flanking sequences of the PAN region revealed that less than 1 kb downstream of PAN is a 2405 bp fragment that is present in M. bovis BCG but absent in the M. bovis wild-type strain. The distribution of the 2405 bp fragment in members of the M. tuberculosis complex was investigated and found to be present in 70 out of 70 M. tuberculosis strains, and 7 out of 7 M. bovis BCG daughter strains, 2 out of 2 Mycobacterium africanum strains, 2 out of 2 Mycobacterium microti strains and 7 out of 25 M. bovis strains. This is the first report of a genetic region of M. bovis BCG that is not universally present in M. bovis strains. The fragment does not appear to be present in any mycobacterial species outside the M. tuberculosis complex. It does not possess any characteristics of known transposable elements and the flanking sequences do not have any obvious features to suggest a deletion mechanism. The genetic location of this region is close to the 3' end of the RD1 region of M. bovis and M. tuberculosis. The polymorphic nature of this locus in M. bovis will provide an additional genetic marker for strain differentiation.
authors.affiliation: Unité de Génétique Mycobactérienne, Institut Pasteur, Paris, France.
authors.forename: Jean
authors.initials: J
authors.lastname: Rauzier
authors.affiliation: Pathobiology Group, Institute of Veterinary, Animal & Biomedical Sciences, Massey University, Palmerston North, New Zealand.
authors.forename: Eamonn
authors.initials: E
authors.lastname: Gormley
authors.affiliation: Laboratoire de Référence des Mycobactéries, Institut Pasteur, paris, France.
authors.forename: M Cristina
authors.initials: MC
authors.lastname: Gutierrez
authors.affiliation: Institut Pasteur, Bangui, Central African Republic.|Unité de Génétique Mycobactérienne, Institut Pasteur, Paris, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Kassa-Kelembho
authors.affiliation: Pathobiology Group, Institute of Veterinary, Animal & Biomedical Sciences, Massey University, Palmerston North, New Zealand.
authors.forename: Laurie J
authors.initials: LJ
authors.lastname: Sandall
authors.affiliation: Pathobiology Group, Institute of Veterinary, Animal & Biomedical Sciences, Massey University, Palmerston North, New Zealand.
authors.forename: Chris
authors.initials: C
authors.lastname: Dupont
authors.affiliation: Unité de Génétique Mycobactérienne, Institut Pasteur, Paris, France.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Gicquel
authors.affiliation: Pathobiology Group, Institute of Veterinary, Animal & Biomedical Sciences, Massey University, Palmerston North, New Zealand.|Unité de Génétique Mycobactérienne, Institut Pasteur, Paris, France.
authors.forename: Alan
authors.initials: A
authors.lastname: Murray
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002417:Cattle; D015342:DNA Probes; D004269:DNA, Bacterial / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D009163:Mycobacterium bovis / Q000145:classification / Q000235:genetics*; D009169:Mycobacterium tuberculosis / Q000145:classification / Q000235:genetics*; D009170:Nontuberculous Mycobacteria / Q000145:classification / Q000235:genetics; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA
chemical_list: D015342:DNA Probes; D004269:DNA, Bacterial