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

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

V. Yu. Kotova, I. V. Manukhov, G. B. Zavilgelskii

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. Key wordsbioluminescence-biosensor-induced promoter-Pcda-PibpA-PgrpE-PkatG-PrecA-PsoxS

Applied Biochemistry and Microbiology 04/2012; 46(8):781-788.

07.04.2012, 2954 просмотра.