Constructing of Bacillus subtilis-Based Lux-Biosensors with the Use of Stress-Inducible Promoters

International Journal of Molecular Sciences

Eugeny Yu. Gnuchikh¹  Vera Yu. Kotova²  Svetlana A. Khrulnova³  Sergey V. Bazhenov³  Eugeniya A. Stepanova³  Uliana S. Novoyatlova³  Andrew G. Kessenikh³  Ilya V. Manukhov³  Anna A. Kudryavtseva³  Maxim V. Bermeshev⁴ 

[1] Kurchatov Genomic Center, State Research Institute of Genetics and Selection of Industrial Microorganisms of the National Research Centre «Kurchatov Institute», 117545 Moscow, Russia;National Research Center «Kurchatov Institute», Kurchatov Complex for Genetic Research, 123098 Moscow, Russia;Research Center for Molecular Mechanisms of Aging and Age-Related Diseases, Moscow Institute of Physics and Technology, 141701 Dolgoprudny, Russia;Topchiev Institute of Petrochemical Synthesis, Russian Academy of Sciences, 119071 Moscow, Russia;

关键词: bacterial biosensor;    Bacillus subtilis;    genotoxicity assessment;    soil analysis;    oxidative stress;    inducible promoters;   

DOI  :  10.3390/ijms22179571

来源: DOAJ

【 摘 要 】

Here, we present a new lux-biosensor based on Bacillus subtilis for detecting of DNA-tropic and oxidative stress-causing agents. Hybrid plasmids pNK-DinC, pNK-AlkA, and pNK-MrgA have been constructed, in which the Photorhabdus luminescens reporter genes luxABCDE are transcribed from the stress-inducible promoters of B. subtilis: the SOS promoter PdinC, the methylation-specific response promoter PalkA, and the oxidative stress promoter PmrgA. The luminescence of B. subtilis-based biosensors specifically increases in response to the appearance in the environment of such common toxicants as mitomycin C, methyl methanesulfonate, and H₂O₂. Comparison with Escherichia coli-based lux-biosensors, where the promoters PdinI, PalkA, and Pdps were used, showed generally similar characteristics. However, for B. subtilis PdinC, a higher response amplitude was observed, and for B. subtilis PalkA, on the contrary, both the amplitude and the range of detectable toxicant concentrations were decreased. B. subtilis PdinC and B. subtilis PmrgA showed increased sensitivity to the genotoxic effects of the 2,2′-bis(bicyclo [2.2.1] heptane) compound, which is a promising propellant, compared to E. coli-based lux-biosensors. The obtained biosensors are applicable for detection of toxicants introduced into soil. Such bacillary biosensors can be used to study the differences in the mechanisms of toxicity against Gram-positive and Gram-negative bacteria.

【 授权许可】

Unknown