Construction of bioluminescent Escherichia coli form lux operon and heat shock promoter for the detection of toxic substances
1999
Yoo, S.O. | Lee, E.K. | Kim, H.S. | Chung, K.H. | Chun, U.H. (Kyung Hee University, Yongin (Korea Republic). Department of Food Technology and Science, Faculty of Life Science)
In order to use heat shock promoter for the detection of toxic substances, dnaK promoter was amplified from E. coli genomic DNA by using a polymerase chain reaction(PCR) followed by suquencing and sub-cloning into the multi-cloning site of the plasmid, pUCD615. The pUCD615 is a broad-host-range vector containing promoterless lux operon orignated from V.fischeri. The recombinant plasmid was transfered to E. coli DH5 alpha through electroporation. The recombinant E. coli showed several patterns of bioluminescent responses to ethanol stress. The bioluminescent E. coli also showed responses to other toxic substances including FeK3(CN)6, CdCl2, rho-nitrophenol and HgCl2. The increases of RLU(Relative Light Unit) were observed at 100 ppm of FeK3(CN)6, 10 ppm and 100 ppm of CdCl2, 1 ppm and 10 ppm of rho-nitrophenol and at 1 ppm of HgCl2.
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