This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Platteeuw, C Articles by de Vos, W M Search for Related Content PubMed PubMed Citation Articles by Platteeuw, C Articles by de Vos, W M Agricola Articles by Platteeuw, C Articles by de Vos, W M

 Previous Article  |  Next Article 

Appl Environ Microbiol. 1994 February; 60(2): 587-593

Use of the Escherichia coli beta-glucuronidase (gusA) gene as a reporter gene for analyzing promoters in lactic acid bacteria.

Department of Biophysical Chemistry, Netherlands Institute for Dairy Research (NIZO), Ede.

ABSTRACT

A transcriptional fusion vector, designated pNZ272, based on the promoterless beta-glucuronidase gene (gusA) of Escherichia coli as a reporter gene, has been constructed for lactic acid bacteria. The replicon of pNZ272 was derived from the Lactococcus lactis plasmid pSH71, allowing replication in a wide range of gram-positive bacteria and E. coli. The applicability of pNZ272 and the expression of the gusA gene in L. lactis was demonstrated in shotgun cloning experiments with lactococcal chromosomal and bacteriophage DNA. In addition, three defined lactococcal promoters were inserted in pNZ272: the plasmid-derived lacA promoter, the chromosomal usp45 promoter, and a promoter from bacteriophage phi SK11G. The three resulting plasmids showed beta-glucuronidase activity in a gusA-deficient E. coli strain and in four species of lactic acid bacteria belonging to the genera Lactobacillus, Lactococcus, and Leuconostoc. The copy numbers of the gusA-expressing plasmids were similar within a single species of lactic acid bacteria. However, the specific beta-glucuronidase activity and the gusA mRNA levels varied considerably both within a single species and among different species of lactic acid bacteria. The transcriptional start site of all three promoters was determined and found to be identical in the different species. The results of this comparative promoter analysis indicate that the requirements for efficient transcription initiation differ among the lactic acid bacteria studied.

This article has been cited by other articles:

• Bachmann, H., Kleerebezem, M., van Hylckama Vlieg, J. E. T. (2008). High-Throughput Identification and Validation of In Situ-Expressed Genes of Lactococcus lactis. Appl. Environ. Microbiol. 74: 4727-4736 [Abstract] [Full Text]  
• Denou, E., Pridmore, R. D., Berger, B., Panoff, J.-M., Arigoni, F., Brussow, H. (2008). Identification of Genes Associated with the Long-Gut-Persistence Phenotype of the Probiotic Lactobacillus johnsonii Strain NCC533 Using a Combination of Genomics and Transcriptome Analysis. J. Bacteriol. 190: 3161-3168 [Abstract] [Full Text]  
• Zahner, D., Scott, J. R. (2008). SipA Is Required for Pilus Formation in Streptococcus pyogenes Serotype M3. J. Bacteriol. 190: 527-535 [Abstract] [Full Text]  
• Cronin, M., Knobel, M., O'Connell-Motherway, M., Fitzgerald, G. F., van Sinderen, D. (2007). Molecular Dissection of a Bifidobacterial Replicon. Appl. Environ. Microbiol. 73: 7858-7866 [Abstract] [Full Text]  
• Arioli, S., Monnet, C., Guglielmetti, S., Parini, C., De Noni, I., Hogenboom, J., Halami, P. M., Mora, D. (2007). Aspartate Biosynthesis Is Essential for the Growth of Streptococcus thermophilus in Milk, and Aspartate Availability Modulates the Level of Urease Activity. Appl. Environ. Microbiol. 73: 5789-5796 [Abstract] [Full Text]  
• Lambert, J. M., Bongers, R. S., Kleerebezem, M. (2007). Cre-lox-Based System for Multiple Gene Deletions and Selectable-Marker Removal in Lactobacillus plantarum. Appl. Environ. Microbiol. 73: 1126-1135 [Abstract] [Full Text]  
• Maze, A., O'Connell-Motherway, M., Fitzgerald, G. F., Deutscher, J., van Sinderen, D. (2007). Identification and Characterization of a Fructose Phosphotransferase System in Bifidobacterium breve UCC2003. Appl. Environ. Microbiol. 73: 545-553 [Abstract] [Full Text]  
• Klijn, A., Moine, D., Delley, M., Mercenier, A., Arigoni, F., Pridmore, R. D. (2006). Construction of a Reporter Vector for the Analysis of Bifidobacterium longum Promoters. Appl. Environ. Microbiol. 72: 7401-7405 [Abstract] [Full Text]  
• Fernandez, M., del Rio, B., Linares, D. M., Martin, M. C., Alvarez, M. A. (2006). Real-time polymerase chain reaction for quantitative detection of histamine-producing bacteria: use in cheese production.. J DAIRY SCI 89: 3763-3769 [Abstract] [Full Text]  
• Bron, P. A., Monk, I. R., Corr, S. C., Hill, C., Gahan, C. G. M. (2006). Novel Luciferase Reporter System for In Vitro and Organ-Specific Monitoring of Differential Gene Expression in Listeria monocytogenes. Appl. Environ. Microbiol. 72: 2876-2884 [Abstract] [Full Text]  
• Ventura, M., Zhang, Z., Cronin, M., Canchaya, C., Kenny, J. G., Fitzgerald, G. F., van Sinderen, D. (2005). The ClgR Protein Regulates Transcription of the clpP Operon in Bifidobacterium breve UCC 2003. J. Bacteriol. 187: 8411-8426 [Abstract] [Full Text]  
• Bongers, R. S., Veening, J.-W., Van Wieringen, M., Kuipers, O. P., Kleerebezem, M. (2005). Development and Characterization of a Subtilin-Regulated Expression System in Bacillus subtilis: Strict Control of Gene Expression by Addition of Subtilin. Appl. Environ. Microbiol. 71: 8818-8824 [Abstract] [Full Text]  
• Pretzer, G., Snel, J., Molenaar, D., Wiersma, A., Bron, P. A., Lambert, J., de Vos, W. M., van der Meer, R., Smits, M. A., Kleerebezem, M. (2005). Biodiversity-Based Identification and Functional Characterization of the Mannose-Specific Adhesin of Lactobacillus plantarum. J. Bacteriol. 187: 6128-6136 [Abstract] [Full Text]  
• van Kranenburg, R., Golic, N., Bongers, R., Leer, R. J., de Vos, W. M., Siezen, R. J., Kleerebezem, M. (2005). Functional Analysis of Three Plasmids from Lactobacillus plantarum. Appl. Environ. Microbiol. 71: 1223-1230 [Abstract] [Full Text]  
• Golic, N., Schliekelmann, M., Fernandez, M., Kleerebezem, M., van Kranenburg, R. (2005). Molecular characterization of the CmbR activator-binding site in the metC-cysK promoter region in Lactococcus lactis. Microbiology 151: 439-446 [Abstract] [Full Text]  
• Barnett, T. C., Patel, A. R., Scott, J. R. (2004). A Novel Sortase, SrtC2, from Streptococcus pyogenes Anchors a Surface Protein Containing a QVPTGV Motif to the Cell Wall. J. Bacteriol. 186: 5865-5875 [Abstract] [Full Text]  
• Verneuil, N., Sanguinetti, M., Le Breton, Y., Posteraro, B., Fadda, G., Auffray, Y., Hartke, A., Giard, J.-C. (2004). Effects of the Enterococcus faecalis hypR Gene Encoding a New Transcriptional Regulator on Oxidative Stress Response and Intracellular Survival within Macrophages. Infect. Immun. 72: 4424-4431 [Abstract] [Full Text]  
• Fernandez, L., Marin, M. L., Langa, S., Martin, R., Reviriego, C., Fernandez, A., Olivares, M., Xaus, J., Rodriguez, J. M. (2004). A Novel Genetic Label for Detection of Specific Probiotic Lactic Acid Bacteria. Food Science and Technology International 10: 101-108 [Abstract]  
• van der Kaaij, H., Desiere, F., Mollet, B., Germond, J.-E. (2004). L-Alanine Auxotrophy of Lactobacillus johnsonii as Demonstrated by Physiological, Genomic, and Gene Complementation Approaches. Appl. Environ. Microbiol. 70: 1869-1873 [Abstract] [Full Text]  
• Bron, P. A., Hoffer, S. M., Van Swam, I. I., De Vos, W. M., Kleerebezem, M. (2004). Selection and Characterization of Conditionally Active Promoters in Lactobacillus plantarum, Using Alanine Racemase as a Promoter Probe. Appl. Environ. Microbiol. 70: 310-317 [Abstract] [Full Text]  
• Pastar, I., Tonic, I., Golic, N., Kojic, M., van Kranenburg, R., Kleerebezem, M., Topisirovic, L., Jovanovic, G. (2003). Identification and Genetic Characterization of a Novel Proteinase, PrtR, from the Human Isolate Lactobacillus rhamnosus BGT10. Appl. Environ. Microbiol. 69: 5802-5811 [Abstract] [Full Text]  
• Varmanen, P., Vogensen, F. K., Hammer, K., Palva, A., Ingmer, H. (2003). ClpE from Lactococcus lactis Promotes Repression of CtsR-Dependent Gene Expression. J. Bacteriol. 185: 5117-5124 [Abstract] [Full Text]  
• Germond, J.-E., Delley, M., Gilbert, C., Atlan, D. (2003). Determination of the Domain of the Lactobacillus delbrueckii subsp. bulgaricus Cell Surface Proteinase PrtB Involved in Attachment to the Cell Wall after Heterologous Expression of the prtB Gene in Lactococcus lactis. Appl. Environ. Microbiol. 69: 3377-3384 [Abstract] [Full Text]  
• Jankovic, I., Ventura, M., Meylan, V., Rouvet, M., Elli, M., Zink, R. (2003). Contribution of Aggregation-Promoting Factor to Maintenance of Cell Shape in Lactobacillusgasseri 4B2. J. Bacteriol. 185: 3288-3296 [Abstract] [Full Text]  
• Neubauer, H., Bauche, A., Mollet, B. (2003). Molecular characterization and expression analysis of the dextransucrase DsrD of Leuconostoc mesenteroides Lcc4 in homologous and heterologous Lactococcus lactis cultures. Microbiology 149: 973-982 [Abstract] [Full Text]  
• Derzelle, S., Hallet, B., Ferain, T., Delcour, J., Hols, P. (2002). Cold Shock Induction of the cspL Gene in Lactobacillus plantarum Involves Transcriptional Regulation. J. Bacteriol. 184: 5518-5523 [Abstract] [Full Text]  
• Bernasconi, E., Germond, J.-E., Delley, M., Fritsche, R., Corthesy, B. (2002). Lactobacillus bulgaricus Proteinase Expressed in Lactococcus lactis Is a Powerful Carrier for Cell Wall-Associated and Secreted Bovine {beta}-Lactoglobulin Fusion Proteins. Appl. Environ. Microbiol. 68: 2917-2923 [Abstract] [Full Text]  
• Solem, C., Jensen, P. R. (2002). Modulation of Gene Expression Made Easy. Appl. Environ. Microbiol. 68: 2397-2403 [Abstract] [Full Text]  
• Barnett, T. C., Scott, J. R. (2002). Differential Recognition of Surface Proteins in Streptococcus pyogenes by Two Sortase Gene Homologs. J. Bacteriol. 184: 2181-2191 [Abstract] [Full Text]  
• Lapierre, L., Mollet, B., Germond, J.-E. (2002). Regulation and Adaptive Evolution of Lactose Operon Expression in Lactobacillus delbrueckii. J. Bacteriol. 184: 928-935 [Abstract] [Full Text]  
• Fernandez, M., Kleerebezem, M., Kuipers, O. P., Siezen, R. J., van Kranenburg, R. (2002). Regulation of the metC-cysK Operon, Involved in Sulfur Metabolism in Lactococcus lactis. J. Bacteriol. 184: 82-90 [Abstract] [Full Text]  
• Thompson, A., Gasson, M. J. (2001). Location Effects of a Reporter Gene on Expression Levels and on Native Protein Synthesis in Lactococcus lactis and Saccharomyces cerevisiae. Appl. Environ. Microbiol. 67: 3434-3439 [Abstract] [Full Text]  
• Vaughan, E. E., van den Bogaard, P. T. C., Catzeddu, P., Kuipers, O. P., de Vos, W. M. (2001). Activation of Silent gal Genes in the lac-gal Regulon of Streptococcus thermophilus. J. Bacteriol. 183: 1184-1194 [Abstract] [Full Text]  
• Gosalbes, M. J., Esteban, C. D., Galán, J. L., Pérez-Martínez, G. (2000). Integrative Food-Grade Expression System Based on the Lactose Regulon of Lactobacillus casei. Appl. Environ. Microbiol. 66: 4822-4828 [Abstract] [Full Text]  
• van den Bogaard, P. T. C., Kleerebezem, M., Kuipers, O. P., de Vos, W. M. (2000). Control of Lactose Transport, beta -Galactosidase Activity, and Glycolysis by CcpA in Streptococcus thermophilus: Evidence for Carbon Catabolite Repression by a Non-Phosphoenolpyruvate-Dependent Phosphotransferase System Sugar. J. Bacteriol. 182: 5982-5989 [Abstract] [Full Text]  
• Stentz, R., Loizel, C., Malleret, C., Zagorec, M. (2000). Development of Genetic Tools for Lactobacillus sakei: Disruption of the beta -Galactosidase Gene and Use of lacZ as a Reporter Gene To Study Regulation of the Putative Copper ATPase, AtkB. Appl. Environ. Microbiol. 66: 4272-4278 [Abstract] [Full Text]  
• Pavan, S., Hols, P., Delcour, J., Geoffroy, M.-C., Grangette, C., Kleerebezem, M., Mercenier, A. (2000). Adaptation of the Nisin-Controlled Expression System in Lactobacillus plantarum: a Tool To Study In Vivo Biological Effects. Appl. Environ. Microbiol. 66: 4427-4432 [Abstract] [Full Text]  
• Giard, J.-C., Rince, A., Capiaux, H., Auffray, Y., Hartke, A. (2000). Inactivation of the Stress- and Starvation-Inducible gls24 Operon Has a Pleiotrophic Effect on Cell Morphology, Stress Sensitivity, and Gene Expression in Enterococcus faecalis. J. Bacteriol. 182: 4512-4520 [Abstract] [Full Text]  
• Ward, D. E., van der Weijden, C. C., van der Merwe, M. J., Westerhoff, H. V., Claiborne, A., Snoep, J. L. (2000). Branched-Chain alpha -Keto Acid Catabolism via the Gene Products of the bkd Operon in Enterococcus faecalis: a New, Secreted Metabolite Serving as a Temporary Redox Sink. J. Bacteriol. 182: 3239-3246 [Abstract] [Full Text]  
• Vriesema, A. J. M., Dankert, J., Zaat, S. A. J. (2000). A Shift from Oral to Blood pH Is a Stimulus for Adaptive Gene Expression of Streptococcus gordonii CH1 and Induces Protection against Oxidative Stress and Enhanced Bacterial Growth by Expression of msrA. Infect. Immun. 68: 1061-1068 [Abstract] [Full Text]  
• Geoffroy, M.-C., Guyard, C., Quatannens, B., Pavan, S., Lange, M., Mercenier, A. (2000). Use of Green Fluorescent Protein To Tag Lactic Acid Bacterium Strains under Development as Live Vaccine Vectors. Appl. Environ. Microbiol. 66: 383-391 [Abstract] [Full Text]  
• Yung, D.-L., McIver, K. S., Scott, J. R., Hollingshead, S. K. (1999). Attenuated Expression of the mga Virulence Regulon in an M Serotype 50 Mouse-Virulent Group A Streptococcal Strain. Infect. Immun. 67: 6691-6694 [Abstract] [Full Text]  
• Looijesteijn, P. J., Boels, I. C., Kleerebezem, M., Hugenholtz, J. (1999). Regulation of Exopolysaccharide Production by Lactococcus lactis subsp. cremoris by the Sugar Source. Appl. Environ. Microbiol. 65: 5003-5008 [Abstract] [Full Text]  
• McCracken, A., Timms, P. (1999). Efficiency of Transcription from Promoter Sequence Variants in Lactobacillus Is Both Strain and Context Dependent. J. Bacteriol. 181: 6569-6572 [Abstract] [Full Text]  
• Ward, D. E., Ross, R. P., van der Weijden, C. C., Snoep, J. L., Claiborne, A. (1999). Catabolism of Branched-Chain alpha -Keto Acids in Enterococcus faecalis: the bkd Gene Cluster, Enzymes, and Metabolic Route. J. Bacteriol. 181: 5433-5442 [Abstract] [Full Text]  
• Gosalbes, M. J., Monedero, V., Pérez-Martínez, G. (1999). Elements Involved in Catabolite Repression and Substrate Induction of the Lactose Operon in Lactobacillus casei. J. Bacteriol. 181: 3928-3934 [Abstract] [Full Text]  
• Brøndsted, L., Hammer, K. (1999). Use of the Integration Elements Encoded by the Temperate Lactococcal Bacteriophage TP901-1 To Obtain Chromosomal Single-Copy Transcriptional Fusions in Lactococcus lactis. Appl. Environ. Microbiol. 65: 752-758 [Abstract] [Full Text]  
• van Kranenburg, R., de Vos, W. M. (1998). Characterization of Multiple Regions Involved in Replication and Mobilization of Plasmid pNZ4000 Coding for Exopolysaccharide Production in Lactococcus lactis. J. Bacteriol. 180: 5285-5290 [Abstract] [Full Text]  
• Vaughan, E. E., Pridmore, R. D., Mollet, B. (1998). Transcriptional Regulation and Evolution of Lactose Genes in the Galactose-Lactose Operon of Lactococcus lactis NCDO2054. J. Bacteriol. 180: 4893-4902 [Abstract] [Full Text]  
• Eichenbaum, Z., Federle, M. J., Marra, D., de Vos, W. M., Kuipers, O. P., Kleerebezem, M., Scott, J. R. (1998). Use of the Lactococcal nisA Promoter To Regulate Gene Expression in Gram-Positive Bacteria: Comparison of Induction Level and Promoter Strength. Appl. Environ. Microbiol. 64: 2763-2769 [Abstract] [Full Text]  
• Meijer, W., van de Bunt, B., Twigt, M., de Jonge, B., Smit, G., Hugenholtz, J. (1998). Lysis of Lactococcus lactis subsp. cremoris SK110 and Its Nisin-Immune Transconjugant in Relation to Flavor Development in Cheese. Appl. Environ. Microbiol. 64: 1950-1953 [Abstract] [Full Text]  
• Hertel, C., Schmidt, G., Fischer, M., Oellers, K., Hammes, W. P. (1998). Oxygen-Dependent Regulation of the Expression of the Catalase Gene katA of Lactobacillus sakei LTH677. Appl. Environ. Microbiol. 64: 1359-1365 [Abstract] [Full Text]  
• Kuipers, O. P., Beerthuyzen, M. M., de Ruyter, P. G. G. A., Luesink, E. J., de Vos, W. M. (1995). Autoregulation of Nisin Biosynthesis in Lactococcus lactis by Signal Transduction. J. Biol. Chem. 270: 27299-27304 [Abstract] [Full Text]