Development of the recombinase-based in vivo expression technology in Streptococcus thermophilus and validation using the lactose operon promoter

Autor: Magali Genay, Maira Junjua, Wessam Galia, Herwig Bachmann, Nadia Gaci, Annie Dary, Yvonne Roussel, Ophélie Uriot, Michiel Kleerebezem
Přispěvatelé: Unité de Recherches Animal et Fonctionnalités des Produits Animaux (URAFPA), Institut National de la Recherche Agronomique (INRA)-Université de Lorraine (UL), NIZO FOOD RESEARCH (NIZO), Nizo food research, Vrije Universiteit Amsterdam [Amsterdam] (VU), Wageningen University and Research Centre [Wageningen] (WUR), Molecular Cell Physiology, AIMMS, Université de Lorraine (UL)-Institut National de la Recherche Agronomique (INRA), NIZO [Ede, Netherlands], Wageningen University and Research [Wageningen] (WUR)
Jazyk: angličtina
Rok vydání: 2014
Předmět:
Streptococcus thermophilus
Operon
[SDV]Life Sciences [q-bio]
lac operon
Applied Microbiology and Biotechnology
Plasmid
LACTIC-ACBACTERIA
HUMAN GUT
Recombinase
Promoter Regions
Genetic
Heat-Shock Proteins
ComputingMilieux_MISCELLANEOUS
GENE-EXPRESSION
lactic-acid bacteria
Lactose operon
0303 health sciences
biology
General Medicine
Genetic Techniques
Lac Operon
proteinase
YOGURT
Plasmids
human gut
Biotechnology
mice
Genetic Vectors
Cre recombinase
system
yogurt
03 medical and health sciences
PROTEINASE
lactococcus-lactis
Animals
Host-Microbe Interactomics
LACTOCOCCUS-LACTIS
Gene
030304 developmental biology
Integrases
IDENTIFICATION
030306 microbiology
transformation
Promoter
biology.organism_classification
gene-expression
Molecular biology
TRANSFORMATION
Mice
Inbred C57BL
MICE
identification
Recombinase-based in vivo expression technology
SYSTEM
Zdroj: Journal of Applied Microbiology
Journal of Applied Microbiology, Wiley, 2014, 116 (3), pp.620-631. ⟨10.1111/jam.12376⟩
Journal of Applied Microbiology, 116(3), 620-631. Wiley-Blackwell
Journal of Applied Microbiology 116 (2014) 3
Journal of Applied Microbiology, 116(3), 620-631
Junjua, M, Galia, W, Gaci, N, Uriot, O, Genay, M, Bachmann, H, Kleerebezem, M, Dary, A & 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, vol. 116, no. 3, pp. 620-631 . https://doi.org/10.1111/jam.12376
ISSN: 1364-5072
1365-2672
Popis: Aims 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. Conclusions 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.
Databáze: OpenAIRE
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