Development of the recombinase-based in vivo expression technology in Streptococcus thermophilus and validation using the lactose operon promoter
Article first published online: 27 NOV 2013
© 2013 The Society for Applied Microbiology
Journal of Applied Microbiology
Volume 116, Issue 3, pages 620–631, March 2014
How to Cite
Junjua, M., Galia, W., Gaci, N., Uriot, O., Genay, M., Bachmann, H., Kleerebezem, M., Dary, A. and Roussel, Y. (2014), Development of the recombinase-based in vivo expression technology in Streptococcus thermophilus and validation using the lactose operon promoter. Journal of Applied Microbiology, 116: 620–631. doi: 10.1111/jam.12376
- Issue published online: 14 FEB 2014
- Article first published online: 27 NOV 2013
- Accepted manuscript online: 29 OCT 2013 02:03AM EST
- Manuscript Accepted: 24 OCT 2013
- Manuscript Revised: 15 OCT 2013
- Manuscript Received: 26 AUG 2013
- HEC (Higher Education Commission, Pakistan)
- University of Lorraine
- Lactose operon;
- recombinase-based in vivo expression technology;
- Streptococcus thermophilus
To construct and validate the recombinase-based in vivo expression technology (R-IVET) tool in Streptococcus thermophilus (ST).
Methods and Results
The R-IVET system we constructed in the LMD-9 strain includes the plasmid pULNcreB allowing transcriptional fusion with the gene of the site-specific recombinase Cre and the chromosomal cassette containing a spectinomycin resistance gene flanked by two loxP sites. When tested in M17 medium, promoters of the genes encoding the protease PrtS, the heat-shock protein Hsp16 and of the lactose operon triggered deletion of the cassette, indicating promoter activity in these conditions. The lactose operon promoter was also found to be activated during the transit in the murine gastrointestinal tract.
The R-IVET system developed in ST is relatively stable, functional, very sensitive and can be used to assay activity of promoters, which are specifically active in in vivo conditions.
Significance and Impact of the Study
This first adaptation of R-IVET to ST provides a highly valuable tool allowing an exploration of the physiological state of ST in the GIT of mammals, fermentation processes or dairy products.