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Monitoring of cell viability under the stress is one of the promising areas of research. The purpose of this work was to study the effect of temperature on the viability of E.coli expressing the mutant firefly luciferases, by measuring the bioluminescence intensity of the endogenous luciferase and the content of intracellular ATP. The object of the study were E. coli BL21 DE3 Codon plus cells transformed with the pETL7 plasmid carrying the firefly luciferase Luciola mingrelica genes: wild-type (WT), thermostable (TS), and green (GTS) mutants. Luciferase was expressed in soluble and active form, and its enzymatic activity was measured by bioluminescence intensity of cells without their destruction on the luminometer (LUM 1, Russia) using a luciferin-based substrate mixture.