title: Comparison of a new insertion element, IS1407, with established molecular markers for the characterization of Mycobacterium celatum.
abstract: Genomic analyses of 18 Mycobacterium celatum strains obtained from different patients in three countries (United States, United Kingdom, and France) were performed; the methods used in this study were restriction fragment length polymorphism (RFLP) analysis, pulsed-field gel electrophoresis (PFGE) analysis, and PCR restriction analysis (PRA) of the hsp-65 gene. A new insertion sequence, IS1407 (GenBank accession no. X97307), belonging to the IS256 family, was identified in M. celatum type 1 strains and was characterized by sequencing. When a probe for Mycobacterium xenopi IS1395-like sequences was used, the RFLP analysis of M. celatum type 1 strains revealed that they contained three or four copies of IS1407 in identical genomic positions, while this element was absent in all M. celatum type 2 strains. PFGE performed with three different endonucleases revealed a unique large restriction fragment (LRF) pattern for M. celatum type 1 strains, whereas the LRF patterns obtained for M. celatum type 2 strains were polymorphic. Moreover, PFGE of nondigested genomic DNA revealed extrachromosomal elements in M. celatum type 2. The type strain of M. celatum type 3 could not be differentiated from M. celatum type 1 strains on the basis of the results of the RFLP analysis, the PFGE analysis, and the PRA of IS1407. In this study we confirmed that M. celatum types 1 and 2 represent distinct genomic clusters and that the molecular markers in M. celatum type 2 exhibit greater polymorphism than the molecular markers in M. celatum type 1.
authors.affiliation: Laboratoire de Référence des Mycobactéries, Institut Pasteur, Paris, France.
authors.forename: M
authors.initials: M
authors.lastname: Picardeau
authors.forename: T J
authors.initials: TJ
authors.lastname: Bull
authors.forename: G
authors.initials: G
authors.lastname: Prod'hom
authors.forename: A L
authors.initials: AL
authors.lastname: Pozniak
authors.forename: D C
authors.initials: DC
authors.lastname: Shanson
authors.forename: V
authors.initials: V
authors.lastname: Vincent
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D018834:Chaperonin 60; D018833:Chaperonins / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis; D016521:Electrophoresis, Gel, Pulsed-Field; D005819:Genetic Markers; D016680:Genome, Bacterial; D006801:Humans; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000235:genetics*; D012150:Polymorphism, Restriction Fragment Length; D015183:Restriction Mapping; D014376:Tuberculosis / Q000382:microbiology*
chemical_list: D001426:Bacterial Proteins; D018834:Chaperonin 60; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D005819:Genetic Markers; C074119:heat-shock protein 65, Mycobacterium; D018833:Chaperonins