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Efficient insertional mutagenesis in lactococci and other gram-positive bacteria
Open accessAbstract
In lactococci, the study of chromosomal genes and their regulation is limited by the lack of an efficient transposon mutagenesis system. We associated the insertion sequence ISS1 with the thermosensitive replicon pG+ host to generate a mutagenic tool that can be used even in poorly transformable strains. ISS1 transposition is random in different lactococcal strains as well as in Enterococcus faecalis and Streptococcus thermophilus. High-frequency random insertion (of about 1%) obtained with this system in Lactococcus lactis allows efficient mutagenesis, with typically one insertion per cell. After ISS1 replicative transposition, the chromosome contains duplicated ISS1 sequences flanking pG+ host. This structure allows cloning of the interrupted gene. In addition, efficient excision of the plasmid leaves a single ISS1 copy at the mutated site, thus generating a stable mutant strain with no foreign markers. Mutants obtained by this transposition system are food grade and can thus be used in fermentation processes.
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Cited by
Structure and Biosynthesis of Two Exopolysaccharides Produced by Lactobacillus johnsonii FI9785
Dertli, E., Colquhoun, I. J., Gunning, A. P., Bongaerts, R. J., Le Gall, G., Bonev, B. B., Mayer, M. J., & Narbad, A. (2013) · Journal of Biological Chemistry · doi:10.1074/jbc.m113.507418 · Cited by 134
Pedersen, M. B., Garrigues, C., Tuphile, K., Brun, C., Vido, K., Bennedsen, M., Møllgaard, H., Gaudu, P., & Gruss, A. (2008) · Journal of Bacteriology · doi:10.1128/jb.00447-08 · Cited by 103
Self-citationPositive role of peptidoglycan breaks in lactococcal biofilm formation
Mercier, C., Durrieu, C., Briandet, R., Domakova, E., Tremblay, J., Buist, G., & Kulakauskas, S. (2002) · Molecular Microbiology · doi:10.1046/j.1365-2958.2002.03160.x · Cited by 88
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Oxaran, V., Ledue-Clier, F., Dieye, Y., Herry, J.-M., Péchoux, C., Meylheuc, T., Briandet, R., Juillard, V., & Piard, J.-C. (2012) · PLoS ONE · doi:10.1371/journal.pone.0050989 · Cited by 73
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Self-citationBurrus, V., Bontemps, C., Decaris, B., & Guédon, G. (2001) · Applied and Environmental Microbiology · doi:10.1128/aem.67.4.1522-1528.2001 · Cited by 62
Lechardeur, D., Fernandez, A., Robert, B., Gaudu, P., Trieu-Cuot, P., Lamberet, G., & Gruss, A. (2010) · Journal of Biological Chemistry · doi:10.1074/jbc.m109.024505 · Cited by 47
Self-citationSaillant, V., Lipuma, D., Ostyn, E., Joubert, L., Boussac, A., Guerin, H., Brandelet, G., Arnoux, P., & Lechardeur, D. (2021) · mBio · doi:10.1128/mbio.03392-20 · Cited by 43
Radziwill-Bienkowska, J. M., Robert, V., Drabot, K., Chain, F., Cherbuy, C., Langella, P., Thomas, M., Bardowski, J. K., Mercier-Bonin, M., & Kowalczyk, M. (2017) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-017-8334-1 · Cited by 30
Metadata
- Authors
- Maguin, E., Prévost, H., Ehrlich, S. D., & Gruss, A.
- Venue
- Journal of Bacteriology
- Publisher
- American Society for Microbiology
- Published
- 1996-02
- Volume
- 178
- Issue
- 3
- Pages
- 931-935
- ISSN
- 0021-9193 (ISSN-L)
- 1098-5530
- OpenAlex
- W2153038986
- Cited by
- 486
- References
- 0
- FWCI
- 4.81
- Type
- journal article
- Language
- en
- Open access
- green
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