A1 Vertaisarvioitu alkuperäisartikkeli tieteellisessä lehdessä
Escherichia coli K-12 (pEGFPluxABCDEamp): a tool for analysis of bacterial killing by antibacterial agents and human complement activities on a real-time basis
Tekijät: Janne Atosuo, Janne Lehtinen, Libor Vojtek, Esa-Matti Lilius
Kustantaja: WILEY-BLACKWELL
Kustannuspaikka: HOBOKEN; 111 RIVER ST, HOBOKEN 07030-5774, NJ USA
Julkaisuvuosi: 2013
Journal: Luminescence
Tietokannassa oleva lehden nimi: Luminescence
Lehden akronyymi: Luminescence
Numero sarjassa: 5
Vuosikerta: 28
Numero: 5
Aloitussivu: 771
Lopetussivu: 779
Sivujen määrä: 9
ISSN: 1522-7235
DOI: https://doi.org/10.1002/bio.2435
Tiivistelmä
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. Copyright (c) 2012 John Wiley & Sons, Ltd.
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. Copyright (c) 2012 John Wiley & Sons, Ltd.
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