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Lux-biosensors for detection of SOS-response, heat shock, and oxidative stress

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Abstract

We have constructed hybrid plasmids pColD and pIbpA in which transcription of reporter genes luxCDABE from Photohabdus luminescens occurred from induced promoters: SOS-promoter of gene cdaA (pColD-CA23) and heat shock gene ibpA (E. coli MG1655 genome), respectively. Main parameters were measured: threshold concentration of an inducer and maximal response and minimal time of response for E. coli lux-biosensors bearing hybrid plasmids pColD and pRecA (SOS-response); pIbpA and pGrpe (heat shock) and pKatG and pSoxS (oxidative stress). Mitomicyn C, cis-diaminedichloroplatinum (SOS-response), ethanol and pentachlorophenol (heat shock), and hydrogen peroxide and methyl viologen (oxidative stress) were used as luminescence inducers. It was shown that the SOS-biosensor pColD is superior in main characteristics to the earlier constructed biosensor pRecA, and the heat biosensor pIbpA is more efficient than the earlier constructed pGrpE. Lux-biosensors (pKatG) and (pSoxS) are high sensitive to hydrogen peroxide and methyl viologen, respectively.

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Abbreviations

AA:

answer amplitude

ROS:

reactive oxygen species

DMSO:

dimethylsulphoxide

4-NCO:

4-nitochinolin-1-oxide

PCR:

polymerase chain reaction

cis-DDP:

cis-diaminedichlorplatine

FMN:

flavinemononucleotid

LB:

Luria-Bertani broth

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Original Russian Text © V.Yu. Kotova, I.V. Manukhov, G.B. Zavilgelskii, 2009, published in Biotekhnologiya, 2009, No. 6, pp. 36–45.

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Kotova, V.Y., Manukhov, I.V. & Zavilgelskii, G.B. Lux-biosensors for detection of SOS-response, heat shock, and oxidative stress. Appl Biochem Microbiol 46, 781–788 (2010). https://doi.org/10.1134/S0003683810080089

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