ATOSUO, Janne, Janne LEHTINEN, Libor VOJTEK and Esa-Matti LILIUS. Escherichia coli K-12 (pEGFPluxABCDEamp): a tool for analysis of bacterial killing by antibacterial agents and human complement activities on a real-time basis. Luminescence. Wiley, 2012, vol. 2012, No 11, p. 1-9. ISSN 1522-7235. Available from: https://dx.doi.org/10.1002/bio.2435.
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Basic information
Original name Escherichia coli K-12 (pEGFPluxABCDEamp): a tool for analysis of bacterial killing by antibacterial agents and human complement activities on a real-time basis
Authors ATOSUO, Janne (246 Finland), Janne LEHTINEN (246 Finland), Libor VOJTEK (203 Czech Republic, belonging to the institution) and Esa-Matti LILIUS (246 Finland, guarantor).
Edition Luminescence, Wiley, 2012, 1522-7235.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 30105 Physiology
Country of publisher Czech Republic
Confidentiality degree is not subject to a state or trade secret
Impact factor Impact factor: 1.273
RIV identification code RIV/00216224:14310/12:00061042
Organization unit Faculty of Science
Doi http://dx.doi.org/10.1002/bio.2435
UT WoS 000328416400025
Keywords in English bioluminescence; bacterial luciferase; Escherichia coli; Photorhabdus luminescens; complement system
Tags AKR, rivok
Tags International impact, Reviewed
Changed by Changed by: Mgr. Libor Vojtek, Ph.D., učo 150465. Changed: 22/3/2013 08:57.
Abstract
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 102–107 colony-forming units (CFU) in E. coli and 103–107 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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