1887

Abstract

An insertion sequence of has been isolated using a trap vector following insertion in and inactivation of the reporter gene. The trap vector is a temperature-sensitive () –mycobacterium shuttle plasmid, pCD4, which contains M, the kanamycin-resistance gene as a selection marker and a expression cassette. The mutation present in pCD4 functions in mycobacteria and enables screening for transposable elements from the mycobacterial genome that disrupt the gene by screening for white colonies on X-Gal plates in both mycobacterial as well as hosts. The vector was used to isolate a novel 1653 kb insertion sequence from named IS. IS duplicated host DNA at the target site, had inverted repeats at its ends and contained two ORFs on one strand. One of the predicted proteins showed homology to a putative transposase from . IS was present in two copies in the genome of . The trap vector appears to be useful in trapping insertion sequences from different mycobacteria by screening for the disrupted LacZ phenotype.

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2000-05-01
2020-01-24
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References

  1. Altschul S. F., Madden T. L., Schaffer A. A., Zhang J., Zhang Z., Miller W., Lipman D. J.. 1997; Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res25:3389–3402[CrossRef]
    [Google Scholar]
  2. Cirillo J. D., Barletta R. G., Bloom B. R., Jacobs W. R. Jr. 1991; A novel transposon trap for mycobacteria: isolation and characterization of IS1096. J Bacteriol173:7772–7780
    [Google Scholar]
  3. Collins D. M., Erasmuson S. K., Stephens D. M., Yates G. F., de Lisle G. W.. 1993; DNA fingerprinting of Mycobacterium bovis strains by restriction fragment analysis and hybridization with insertion elements IS1091 and IS6110. J Clin Microbiol31:1143–1147
    [Google Scholar]
  4. Connell N. D.. 1994; Mycobacterium: isolation, maintenance, transformation and mutant selection. Methods Cell Biol45:107–125
    [Google Scholar]
  5. Dellagostin O. A., Wall S., Norman E., O’Shaughnessy T., Dale J. W., McFadden J.. 1993; Construction and use of integrative vectors to express foreign genes in mycobacteria. Mol Microbiol10:983–993[CrossRef]
    [Google Scholar]
  6. England P. M., Wall S., McFadden J. J.. 1991; IS900 promoted stable integration of a foreign gene into mycobacterium. Mol Microbiol5:2047–2052[CrossRef]
    [Google Scholar]
  7. Guilhot C., Gicquel B., Davis J., Martin C.. 1992; Isolation and analysis of IS6120, a new insertion sequence from Mycobacteriun smegmatis. Mol Microbiol6:107–113[CrossRef]
    [Google Scholar]
  8. Guilhot C., Otal I., van Rompney I., Martin C., Gicquel B.. 1994; Efficient transposition in mycobacteria: construction of Mycobacterium smegmatis insertional mutant libraries. J Bacteriol176:535–539
    [Google Scholar]
  9. Holland K. T., Ratledge C.. 1971; A procedure for selecting and isolating specific auxotrophic mutants of Mycobacterium smegmatis. J Gen Microbiol66:115–118[CrossRef]
    [Google Scholar]
  10. Kumar D., Srivastava R., Srivastava B. S.. 1998; Genetic rearrangements leading to disruption of heterologous gene expression in mycobacteria: an observation with E. coli β-galactosidase in Mycobacterium smegmatis and its implication in vaccine development. Vaccine16:1212–1215[CrossRef]
    [Google Scholar]
  11. Kunze Z. M., Wall S., Appleberg R., Silva M. T., Partaels F., McFadden J.. 1991; IS901, a new member of a widespread class of atypical insertion sequences is associated with pathogenicity in Mycobacterium avium. Mol Microbiol5:2265–2272[CrossRef]
    [Google Scholar]
  12. McAdam R. A., Guilhot C., Gicquel B.. 1994; Transposition in mycobacteria. In Tuberculosis: Pathogenesis, Protection and Control pp.199–216Edited by Bloom B. R.. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  13. McFadden J. J., Butcher P. D., Chiodini R. J., Hermon-Taylor J.. 1987; Use of DNA probes to distinguish between mycobacterial species: Crohn’s disease-isolated mycobacteria are identical to Mycobacterium paratuberculosis. J Clin Microbiol25:796–801
    [Google Scholar]
  14. Mahillon J., Chandler M.. 1998; Insertion sequences. Microbiol Mol Biol Rev62:725–774
    [Google Scholar]
  15. Martin C., Timm J., Rauzier J., Gomez-Lus R., Davis J., Gicquel B.. 1990; Transposition of an antibiotic resistance element in mycobacteria. Nature345:739–742[CrossRef]
    [Google Scholar]
  16. Mazurek G. H., Cave M. D., Eisenach K. D., Wallace R. J. Jr, Bates J. H., Crawford J. T.. 1991; Chromosomal DNA fingerprint patterns produced with IS6110 as strain-specific markers for epidemiologic studies of tuberculosis. J Clin Microbiol29:2030–2033
    [Google Scholar]
  17. Pelicic V., Jackson M., Reynat J.-M., Jacob W. R. Jr, Gicquel B., Guilhot C.. 1997; Efficient allelic exchange and transposon mutagenesis in Mycobacterium tuberculosis. Proc Natl Acad Sci USA94:10955–10960[CrossRef]
    [Google Scholar]
  18. Sambrook J., Fritsch E. F., Maniatis T.. 1989; Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  19. Silcox V. A., Good R. A., Flyod M. M.. 1981; Identification of clinically significant Mycobacterium fortuitum complex isolates. J Clin Microbiol13:908–912
    [Google Scholar]
  20. Small P. M., Van Embden J. D. A.. 1994; Molecular epidemiology of tuberculosis. In Tuberculosis: Pathogenesis, Protection and Control pp.569–582Edited by Bloom B. R. . Washington, DC: American Society for Microbiology;
    [Google Scholar]
  21. Stover C. K., de la Cruz V. F., Fuerst T. R..11 other authors 1991; New use of BCG for recombinant vaccines. Nature351:456–460[CrossRef]
    [Google Scholar]
  22. Takemura H., Horinouchi S., Beppu T.. 1991; Novel insertion sequence IS1380 from Acetobacter pasteurianus is involved in loss of ethanol-oxidizing ability. J Bacteriol173:7070–7076
    [Google Scholar]
  23. Thierry D., Brisson-Noel A., Vincent-Levy-Frebault V., Nguyen S., Guesdon J. L., Gicquel B.. 1990; Characterization of a Mycobacterium tuberculosis insertion sequence, IS6110, and its application in diagnosis. J Clin Microbiol28:2668–2673
    [Google Scholar]
  24. Wallace R. J. Jr, Brown B. A., Silcox V. A..8 other authors 1991; Clinical disease, drug susceptibility and biochemical patterns of the unnamed third biovariant complex of M. fortuitum. J Infect Dis163:598–603[CrossRef]
    [Google Scholar]
  25. Woods G. L., Washington J. A.. 1987; Mycobacteria other than Mycobacterium tuberculosis: review of microbiologic and clinical aspects. Rev Infect Dis9:275–294[CrossRef]
    [Google Scholar]
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