Characterization of In Vivo Reporter Systems for Gene Expression and Biosensor Applications Based on luxAB Luciferase Genes

Blouin, Kim; Walker, Stephen G.; Smit, J; Turner, Rufus

doi:10.1128/aem.62.6.2013-2021.1996

Cited by 37 publications

(15 citation statements)

References 50 publications

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“…The same has been reported when luxABCDE genes from P. luminescens were integrated into the E. coli chromosome . The availability of fatty aldehyde has been reported to be the limiting factor in P. luminescens luciferase‐linked BL emission . We also added a fatty aldehyde (decanal) into a growth medium for recombinant E. coli –lux and observed that the BL signal increased to the level indicated by the increase of the OD in the logarithmic phase (data not shown).…”

Section: Discussionsupporting

confidence: 72%

“…We have previously used luxAB genes from V. harveyi in E. coli . The problem with this construct was that during the logarithmic growth phase the BL signal elevated considerably more than indicated by OD signal and at the beginning of the stationary phase BL emission collapsed.…”

Section: Discussionmentioning

confidence: 99%

“…Another method for measuring the number of bacterial cells is the use of bacteria with reporter genes, encoding products with detectable functionality linked to the viability of the cells, such as insect luciferase ( lucFF ) from the firefly Photinus pyralis or bacterial luciferase ( luxAB ) from Vibrio harveyi . Expression of these genes results in bacterial cells emitting bioluminescence (BL) . Green fluorescent protein (GFP) is another commonly used reporter.…”

Section: Introductionmentioning

confidence: 99%

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Escherichia coli K‐12 (pEGFPluxABCDEamp): a tool for analysis of bacterial killing by antibacterial agents and human complement activities on a real‐time basis

et al. 2012

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Photorhabdus luminescens luxCDABE genes were integrated into E. coli K-12 using a high copy number plasmid containing modified luxABCDE genes under the control of the powerful Lac promoter. This strain emitted 10 times higher bioluminescence (BL) than P. luminescens. BL production under different growth conditions was studied. In both bacterial strains, the increase in BL signal correlated with the increase in optical density (OD) in a rich growth medium. However, at the logarithmic growth phase, the BL signal was roughly constant. By contrast, in minimal growth media, there was no substantial growth and the BL/cell was approximately five times higher than in the rich medium. The dynamic measurement range of BL was 10(2) -10(7) colony-forming units (CFU) in E. coli and 10(3) -10(7) CFU in P. luminescens. Because the decrease in the BL signal correlated with the decrease in CFU and OD, i.e. the number of bacterial cells killed, it proved to be very suitable for assessing the antibacterial effects of different antimicrobial agents. Unlike with plate counting, the kinetics of killing can be monitored on a real-time basis using BL measurements. Complement activities in different samples can be estimated using only one serum dilution. The transformed E. coli strain appeared to be superior to P. luminescens in these applications because E. coli was complement sensitive, the detection limit of E. coli was one order lower and the BL-producing system of P. luminescens appeared to be quite unstable.

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Section: Discussionsupporting

confidence: 72%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Escherichia coli K‐12 (pEGFPluxABCDEamp): a tool for analysis of bacterial killing by antibacterial agents and human complement activities on a real‐time basis

et al. 2012

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“…We decided to maintain the pH, oxygen and temperature constant and to study the influence of the bioluminescent strains, their growth stage and growth medium on the spectral emission. Although tetradecanal is the natural aldehyde in the natural bioluminescence reaction (27), decanal was supplied in our experiments as previously mentioned for its practical use (28). From our results, there was no difference in the spectral emission between natural or recombinant cells or between Gram-positive and Gram-negative strains.…”

Section: Discussionmentioning

confidence: 56%

Comparison of the spectral emission of lux recombinant and bioluminescent marine bacteria

et al. 2003

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The purpose of the present paper was to study the influence of bacteria harbouring the luciferase-encoding Vibrio harveyi luxAB genes upon the spectral emission during growth in batch-culture conditions. In vivo bioluminescence spectra were compared from several bioluminescent strains, either naturally luminescent (Vibrio fischeri and Vibrio harveyi) or in recombinant strains (two Gram-negative Escherichia coli::luxAB strains and a Gram-positive Bacillus subtilis::luxAB strain). Spectral emission was recorded from 400 nm to 750 nm using a highly sensitive spectrometer initially devoted to Raman scattering. Two peaks were clearly identified, one at 491-500 nm (+/- 5 nm) and a second peak at 585-595 (+/- 5 nm) with the Raman CCD. The former peak was the only one detected with traditional spectrometers with a photomultiplier detector commonly used for spectral emission measurement, due to their lack of sensitivity and low resolution in the 550-650 nm window. When spectra were compared between all the studied bacteria, no difference was observed between natural or recombinant cells, between Gram-positive and Gram-negative strains, and growth conditions and growth medium were not found to modify the spectrum of light emission.

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“…All five RR‐dependent promoters‐ lux (luciferase) fusions were inserted into pLUX vector (Table ) (Blouin et al . ; Burton et al . ).…”

Section: Methodsmentioning

confidence: 99%

Cross talk in promoter recognition between six NarL‐family response regulators of Escherichia coli two‐component system

Yoshida

Ishihama

2015

Genes to Cells

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Bacterial two‐component system (TCS) is composed of the sensor kinase (SK) and the response regulator (RR). After monitoring an environmental signal or condition, SK activates RR through phosphorylation, ultimately leading to the signal‐dependent regulation of genome transcription. In Escherichia coli, a total of more than 30 SK‐RR pairs exist, each forming a cognate signal transduction system. Cross talk of the signal transduction takes place at three stages: signal recognition by SK (stage 1); RR phosphorylation by SK (stage 2); and target recognition by RR (stage 3). Previously, we analyzed the stage 2 cross talk between the whole set of E. coli SK‐RR pairs and found that the cross talk takes place for certain combinations. As an initial attempt to identify the stage 3 cross talk at the step of target promoter recognition by RR, we analyzed in this study the cross‐recognition of target promoters by six NarL‐family RRs, EvgA, NarL, NarP, RcsB, UhpA, and UvrY. Results of both in vivo and in vitro studies indicated that the stage 3 cross talk takes place for limited combinations, in particular, including a multifactor‐regulated ydeP promoter.

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Characterization of In Vivo Reporter Systems for Gene Expression and Biosensor Applications Based on luxAB Luciferase Genes

Cited by 37 publications

References 50 publications

Escherichia coli K‐12 (pEGFPluxABCDEamp): a tool for analysis of bacterial killing by antibacterial agents and human complement activities on a real‐time basis

Escherichia coli K‐12 (pEGFPluxABCDEamp): a tool for analysis of bacterial killing by antibacterial agents and human complement activities on a real‐time basis

Comparison of the spectral emission of lux recombinant and bioluminescent marine bacteria

Cross talk in promoter recognition between six NarL‐family response regulators of Escherichia coli two‐component system

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