NOVEL METHOD FOR DETECTING AND QUANTITATING TARGET ENZYME ACTIVITY USING ARTIFICIAL GENETIC CIRCUITRY

摘要

PURPOSE: A novel method for detecting and quantitating of target enzyme activation using genetic enzyme screening system(GESS) is provided to sense useful gene in various gene groups and to be used in protein engineering. CONSTITUTION: A genetic enzyme screening system(GESS) for sensing phenol compounds contains a gene which encodes regulatory protein inducing reporter protein expression of downstream and recognizes DmpR(transcriptional regulator for dimethylphenol), XylR(transcriptional regulator for xylene), MopR(transcriptional regulator for phenol metabolism), PhhR(transcriptional regulator for phenol and monomethylated phenols metabolism), PhlR(transcriptional regulator for methylphenol degradation), or TbuT(transcriptional regulator for toluene-benzene utilization) phenol compounds; one or more reporter genes selected from the group consisting of a gene encoding fluorescent protein and an antibiotic resistance gene; and a gene expression regulatory site. The reporter gene is interconnected with a promoter which regulates reporter gene expression.