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Conjugating DNA into Mycobacteria

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Mycobacteria Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 101))

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Abstract

Bacterial conjugation is a phenomenon that is more widespread than was originally thought. It medtates genetrc exchange not only between cells of the same species, but also between a wide variety of unrelated organisms. Conjugation between gram-negative and gram-positive bacteria was first demonstrated using recombinant-shuttle plasmids (1). In Mycobacterium, the shuttle-plasmid pMY10 was introduced to Mycobacterium smegmatis from Escherichia coli (2), as was the naturally occurring plasmid RSF1010 (3). More dramatic examples of conjugation have been shown between E. coli and yeast cells and from Agrobacterium spp. to tobacco-plant cells (4,5). This would suggest that in the horizontal transfer of genetic material, few barriers exist that prevent the acquisition of new genetic information, rather the ability of the DNA to be maintained in the new host would seem to be the main barrier to be overcome.

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References

  1. Trieu-Cuot, P, Carlier, C., Martin, P., and Courvalin, P. (1987) Plasmid transfer by conjugation from Escherichia coli to Gram-positive bacteria. FEMS Microbiol Lett. 48, 289–294

    Article  CAS  Google Scholar 

  2. Lazraq, R., Clavel-Seres, S, David, H L., and Rolland-Dussoix, D. (1990) Conjugative transfer of a shuttle plasmid from Escherichia coli to Mycobacterium smegmatis FEMS. Microbial Lett. 69, 135–138

    Article  CAS  Google Scholar 

  3. Gormley, E. P. and Davies,.I. (1991) Transfer of plasmid RSF1010 by conmgatron from Escherichia coli to Streptomyces lividans and Mycobacterium smegmatis. J. Bacteriol 173, 6705–6708.

    CAS  PubMed  Google Scholar 

  4. Buchanan-Wollaston, V., Passiatore, J. E., and Cannon, F. (1987) The mob and oriT mobilization functions of a bacterial plasmid promote its transfer to plants. Nature 328, 172–175

    Article  CAS  Google Scholar 

  5. Heinemann, J A and Sprague, Jr, G F (1989) Bacterial conjugative plasmids mobilize DNA transfer between bacteria and yeast. Nature 340, 205–209.

    Article  CAS  PubMed  Google Scholar 

  6. Simon, R, Priefer, U, and Puhler, A (1983) A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gram-negative bacteria Bio/Technology 1, 784–791

    Article  CAS  Google Scholar 

  7. Herrero, M, De Lorenzo, V., and Timmis, K N. (1990) Transposon vectors containing non-antibiotic resistance selection markers for cloning and stable chromosomal insertion of foreign genes in Gram-negative bacteria. J Bacteriol 172, 6557–6567

    CAS  PubMed  Google Scholar 

  8. Guilhot, C., Gicquel, B, and Martin, C. (1992) Temperature-sensitive mutants of the Mycobacterium plasmid pAL5000 FEMS Microbiol Lett. 98, 181–186

    Article  CAS  Google Scholar 

  9. Guilhot, C., Otal, I., Van Rompaey, I, Martin, C and Gicquel, B.(1994) Efficient transposition in mycobacteria construction of Mycobacterium smegmatis insertional mutant libraries. J Bacteriol 176, 535–539

    CAS  PubMed  Google Scholar 

  10. Gavigan, J A, Guilhot, C., Gicquel, B. and Martin, C. (1995) Use of conJugative and thermosensitive cloning vectors for transposon delivery to Mycobacterium smegmatis FEMS Microbiol 127, 35–39

    Article  CAS  Google Scholar 

  11. Snapper, S. B., Melton, R E, Mustafa, S, Kieser, T, and Jacobs, Jr, W R. (1990) Isolation and characterisation of efficient transformation mutants of Mycobacterium smegmatis. Mel Mlcroblol 4, 1911–1919

    Article  CAS  Google Scholar 

  12. Hermans, J, Boschloo, J G, and de Bont, JAM. (1990) Transformation of Mycobacterium aurum by electroporation. the use of glycine, lysozyme and isonicotinic acid hydrazide in enhancing transformation efficiency FEMS Microbiol. Lett. 72, 221–224

    Article  CAS  Google Scholar 

  13. Gauigan, J A., Ainsa, J. A., Perez, E, Otal, I, and Martin, C (1997) J Bactriol 179, 4115–4122

    Google Scholar 

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Author information

Authors and Affiliations

  1. Departmento de Microbiologia, Facultad de Medicina, Universidad de Zaragoza, Zaragoza, Spain

    Carlos Martín

Authors
  1. Julie-Ann Gavigan
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  2. Carlos Martín
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Editor information

Editors and Affiliations

  1. London School of Hygiene and Tropical Medicine, London, UK

    Tanya Parish  & Neil G. Stoker  & 

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© 1998 Humana Press Inc., Totowa, NJ

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Gavigan, JA., Martín, C. (1998). Conjugating DNA into Mycobacteria. In: Parish, T., Stoker, N.G. (eds) Mycobacteria Protocols. Methods in Molecular Biology™, vol 101. Humana Press. https://doi.org/10.1385/0-89603-471-2:119

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  • DOI: https://doi.org/10.1385/0-89603-471-2:119

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-471-6

  • Online ISBN: 978-1-59259-576-1

  • eBook Packages: Springer Protocols

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