ASSAY OF ATP IN INTACT ESCHERICHIA-COLI-CELLS EXPRESSING RECOMBINANT FIREFLY LUCIFERASE

Properties of recombinant firefly luciferase synthesized by Escherichi a coli cells inside the intact cells were studied. Low concentrations of polymyxin B was used to increase the permeability of E. coli cell m embranes, allowing the concentration of the luciferase substrates insi de the cells to be varied. The dependence of bioluminescence intensity on intracellular ATP concentration was determined. Recombinant cells carrying the firefly luciferase gene can be used for fast screening of substances that influence the synthesis and hydrolysis of ATP inside the cells, as well as transport of substrates through the cell membran e.