METHOD FOR SCREENING OF USEFUL ENZYMES FROM METAGENOME USING FORCED TRANSCRIPTION

摘要

PURPOSE: A high efficiency detection method of metagenome-derived useful enzyme is provided to enhance the expression of a foreign gene using a forces transcription system, to sense a small amount of reactant, and to sort a target gene. CONSTITUTION: A high efficiency detection method for metagenome-derived enzyme through forced transcription comprises: a step of randomly inserting transposon into a metagenome library gene containing a gene encoding an enzyme to be detected; a step of transducing the metagenome library to a RNA polymerase-inserted host microbe and preparing a recombinant microbe library; a step of introducing a redesigned genetic circuit for sensing phenol compounds to the metagenome library; a step of treating an inducer to the recombinant microbe library and culturing; a step of treating the recombinant microbe library with a phenol releasing compound; a step of measuring and detecting activity of a reporter protein at high speed; and a step of collecting a gene of the enzyme which releases the phenol compounds from a detected microbe and performing sequencing of the gene. [Reference numerals] (AA) Redesigned genetic circuit searching system; (BB) Redesigned genetic circuit; (CC) Forced transcription introducing library; (DD) Forced transcription system; (EE) Metagenome library; (FF) Forced transcription unit;