Abstract
Reporter gene is a generic term that is used to describe genes whose expression is indicative of the presence of a particular cell, a specific event occurring within that cell, or a reflection of the expression of other, usually targeted genes of interest. For example, the presence of such a gene may make an organism standout to help in identification, as will be described in this chapter, or it could be that a gene is only expressed under certain conditions, e.g., during different developmental stages (1), environmental stresses (2,3) , or DNA repair as a result of induction of the SOS response to DNA damage (4). Naturally-occurring reporter genes can be used to avoid the need for genetic engineering by selecting a unique property of an organism and using it as the reporter. However, by far the commonest approach is to introduce a reporter gene into a cell to allow detection of a particular organism or to monitor its activity. The reporter gene can be located on a plasmid or to increase the stability of the gene within the cell it can be incorporated into the bacterial chromosome. There are a wide variety of reporter genes available, and all have particular properties that make them more or less useful depending on the application (Table 1). The choice of a particular reporter gene is made on the basis of the environment into which the organism is to be released, how the gene is to be detected, and what factors might interfere with the detection of the gene and/or its protein product.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Srikantha, T., Klapach, A., Lorenz, W. W., Tsai, L. K., Laughlin, L. A., Gorman, J. A., and Soll, D. R. (1996) The sea pansy Renilla reniformis luciferase serves as a sensitive bioluminescent reporter for differential gene expression in Candida albicans. J. Bact. 178, 121–129.
Schrogel, O. and R. Allmansberger (1997) Optimization of the BgaB reporter system: Determination of transcriptional regulation of stress responsive genes in Bacillus subtilis. FEMS Microbiol. Lett. 153, 237–243.
Kragelund, L., Hosbond, C., and Nybroe, O. (1997) Distribution of metabolic activity and phosphate starvation response of lux-tagged Pseudomonas fluorescens reporter bacteria in barley rhizosphere. Appl. Environ. Microbiol. 63, 4920–4928.
Volmer, A. C., Belkin, S., Smulski, D. R., Van Dyk, T.K., and La Rossa, R.A. (1997) Detection of DNA damage by use of Escherichia coli Carrying recA′-lux, uvrA′-lux or alkA′-lux reporter plasmids. Appl. Environ. Microbiol. 63, 2566–2571.
Bale, M. J., Fry, J. C., and M. J. Day (1987) Plasmid transfer between strains of Pseudomonas aeruginosa on membrane filters attached to river stones. J. Gen. Microbiol. 133, 3099–3107.
Ramos, J. L., Duque, E., and Ramos-Gonzalez, M.-I. (1991) Survival in soils of an herbicide-resistant Pseudomonas putida strain bearing a recombinant TOL plasmid. Appl. Environ. Microbiol. 57, 260–266.
Hofte, M., Mergeay, M., and Verstraete, W. (1990) Marking the Rhizopseudomonas strain 7NSK2 with a mu d(lac) element for ecological studies. Appl. Environ. Microbiol. 56, 1046–1052.
Drahos, D. J., Barry, G.F., Hamming, B.C., Brant, E. L., Kilne, E. L., and Skipper, H. D. et al. (1992) Spread and survival of genetically marked bacteria in soil, in Release of Genetically Engineered and Other Micro-organisms, (Fry, J. C., and Day, M. J., eds.) Cambridge University Press, Cambridge, UK, pp. 147–159.
Robinson, P. J., Walker, J. T., Keevil, C. W., and Cole, J. (1995) Reporter genes and fluorescent-probes for studying the colonization of biofilms in a drinking-water supply line by enteric bacteria. FEMS Microbiol. Lett. 129, 183–188.
Wipat, A., Wellington, E. M. H., and Saunders, V. A. (1991) Streptomyces marker plasmids For monitoring survival and spread of streptomycetes in soil. Appl. Environ. Microbiol. 57, 3322–3330.
Winstanley, C., Morgan, J. A. W., Pickup, R. W., Jones, J. G., and Saunders, J. R. (1989) Differential regulation of lambda pL and pR promoters by a cI repressor in a broad-host-range thermoregulated plasmid marker system. Appl. Environ. Micro. 55, 771–777.
Morgan, J. A. W., Winstanley, C., Pickup, R. W., Jones, J. G., and Saunders, J. R. (1989) Direct phenotypic and genotypic detection of a recombinant pseudomonad population released into lake water. Appl. Environ. Microbiol. 55, 2537–2544.
Winstanley, C., Morgan, J. A. W., Pickup, R. W., Jones, J. G., and Saunders, J. R. (1991) Use of a xylE marker gene to monitor survival of recombinant Pseudomonas putida populations in lake water by culture on nonselective media. Appl. Environ. Microbiol. 57, 1905–1913.
Morgan, J. A. W., Cranwell, P. A., and Pickup, R. W. (1991) Survival of Aeromonas salmonicida in lake water. Appl. Environ. Microbiol. 57, 1777–1782.
Burlage, R. S., Yang, Z. K., and Mehlhorn, T. (1996) A transposon for green fluorescent protein transcriptional fusions: Application for bacterial transport experiments. Gene 173, 53–58.
Josenhans, C., Friedrich, S., and Suerbaum, S. (1998) green fluorescent protein as a novel marker and reporter system in Helicobacter sp. FEMS Microbiol. Lett. 161, 263–273.
Suarez, A., Güttler, A., Strätz, M., Staendner, L. H., Timmis, K. N., and Guzmán, C. A. (1997) Green fluorescent protein-based reporter systems for genetic analysis of bacteria including monocopy applications. Gene 196, 69–74.
Lybarger, L., Dempsey, D., Patterson, G. H., Piston, D. W., Kain, S. R., and Chervenak, R. (1998) Dual-color flow cytometric detection of fluorescent proteins using single-laser (488-nm) excitation. Cytometry. 31, 147–152.
Andersen, J. B., Sternberg, C., Poulsen, L. K., Bjorn, S. P., Givskov, M., and Molin, S. (1998) New unstable variants of green fluorescent protein for studies of transient gene expression in bacteria. Appl. Environ. Microbiol. 64, 2240–2246.
Rattray. E. A. S., Prosser, J. I., Killham, K., and Glover, L. A. (1990) Luminescence-based non-extractive technique for in situ detection of E. coli in soil. Appl. Environ. Microbiol. 56, 3368–3374.
Rattray. E. A. S., Tyrrell, J. A., Prosser, J. I., Glover, L. A., and Killham, K. (1993) Effect of bulk density and temperature on wheat rhizosphere colonization by lux-modified Pseudomonas fluorescens. Eur. J. Soil Biol. 29, 73–82.
Prosser, J. I., Killham, K., Glover, L. A., and Rattray, E. A. S. (1996) Luminescence-based systems for detection of bacteria in the environment. Crit. Rev. Biotech. 16, 157–183.
Pearson, R. E., Jurgensen, S., Sarkis, G. J., Hatfull, G. F., and Jacobs, W. R. (1996) Construction of D29 shuttle plasmids and luciferase reporter phages for detection of Mycobacteria. Gene. 183, 129–136.
Carriere, Riska, C. P. F., Zimhony, O., Kriakov, J., Bardarov, S., Burns, J., et al. (1997) Conditionally replicating luciferase reporter phages: Improved sensitivity for rapid detection and assessment of drug susceptibility of Mycobacterium tuberculosis. J. Clin. Microbiol. 35, 3232–3239.
Loessner, M. J., Rees, C. E. D., Stewart, G. S. A. B., and Scherer, S. (1996) Construction of luciferase reporter bacteriophage A511::luxAB for rapid and sensitive detection of viable listeria cells. Appl. Environ. Microbiol. 62, 1133–1140.
Applegate, B., Kelly, C., Lackey, L., McPherson, J., Kehrmeyer, S., Menn, F. M., et al. (1997) Pseudomonas putida B2: A tod-lux bioluminescent reporter for toluene and trichloroethylene co-metabolism. J. Indust. Microbiol. Biotech. 18, 4–9.
Heitzer, A., Applegate, B., Kehrmeyer, S., Pinkart, H., Webb, O. F., Phelps, T. J., et al. (1998) Physiological considerations of environmental applications of lux reporter fusions. J. Microbiol. Meth. 33, 45–57.
Blatny. J. M., Brautaset, T., Winther-Larsen, H. C., Haugan, K., and Valla, S. (1997) Construction and use of a versatile set of broad-host-range cloning and expression vectors based on the RK2 replicon. Appl. Environ. Microbiol. 63, 370–379.
Blatny. J. M., Brautaset, T., Winther-Larsen, H. C., Karunakaran, P., and Valla, S. (1997) Improved broad-host-range RK2 vectors useful for high and low regulated gene expression levels in Gram-negative bacteria. Plasmid. 38, 35–51.
Baulcombe, D. C., Chapman, S., and Cruz, S. S. (1995) Jellyfish green fluorescent protein as a reporter for virus-infections. Plant. J. 7, 1045–1053.
Liu, Y., Golden, S. S., Kondo, T., Ishiura, M., and Johnson, C. H. (1995) Bacterial luciferase as a reporter of circadian gene-expression in cyanobacteria. J. Bact. 177, 2080–2086.
Flaherty, J. E., Weaver, M. A., Payne, G. A., and Woloshuk, C. P. (1995) A beta-glucuronidase reporter gene construct for monitoring aflatoxin biosynthesis in Aspergillus flavus. Appl. Environ. Microbiol. 61, 2482–2486.
Bullifent, H. L., Moir, A., and Titball, R. W. (1995) The construction of a reporter system and use for the investigation of Clostridium perfringens gene-expression. Microbiol. Lett. 131, 99–105.
Arain, T. M., Resconi, A. E., Singh, D. C., and Stover, C. K. (1996) Reporter gene technology to assess activity of antimycobacterial agents in macrophages. Antimic. Agents and Chemo. 40, 1542–1544.
Bundy, J. G., Wardell, J. L., Campbell, C. D., Killham, K., and Paton, G. I. (1997) Application of bioluminescence-based microbial biosensors to the ecotoxicity of organotins. Lett. Appl. Microbiol. 25, 353–358.
Ulijasz, A. T., Grenader, A., and Weisblum, B. (1996) A vancomycin-inducible lacZ reporter system in Bacillus subtilis: Induction by antibiotics that inhibit cell wall synthesis and by lysozyme. J. Bact. 178, 6305–6309.
Chang, J. R., and Geider, K. (1995) The Use of luciferase as a reporter for response of plant-cells to the fireblight pathogen Erwinia amylovora. Plant Cells Rep. 14, 497–500.
Simmons, J. S. (1926) A culture medium for differentiating organisms of typhoidcolon aerogenes groups and for isolation of certain fungi. J. Infect. Dis. 39, 209.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1999 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Firth, J.R. (1999). Reporter Gene Expression for Monitoring Microorganisms in the Environment. In: Edwards, C. (eds) Environmental Monitoring of Bacteria. Methods in Biotechnology, vol 12. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-566-2:187
Download citation
DOI: https://doi.org/10.1385/0-89603-566-2:187
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-0-89603-566-9
Online ISBN: 978-1-59259-487-0
eBook Packages: Springer Protocols