Patent ID: 6297013
Filing Date: 2001-10-02
Classification: C12N

Abstract:
A method for detecting the presence of a sequence-specific DNA-binding molecule capable of specifically binding to a nucleotide sequence, comprising:(a) providing:(i) a first sample suspected of containing a sequence-specific DNA-binding molecule capable of specifically binding to a first nucleotide sequence;(ii) a restriction enzyme capable of binding to a second nucleotide sequence and cleaving at a restriction site; and(iii) a recombinant plasmid comprising said first and second nucleotide sequences and said restriction site, wherein said restriction site is located at a position selected from being within said first nucleotide sequence, within said second nucleotide sequence, substantially adjacent to said first nucleotide sequence, and substantially adjacent to said second nucleotide sequence; and(b) contacting said first sample with said recombinant plasmid to generate a contacted sample, under conditions such that said sequence-specific DNA-binding molecule specifically binds to said first nucleotide sequence in said recombinant plasmid to generate a bound plasmid;(c) treating said contacted sample with said restriction enzyme under conditions such that said bound plasmid is not substantially cleaved by said restriction enzyme; and(d) detecting the presence of uncleaved plasmid in said contacted sample, thereby detecting the presence of said sequence-specific DNA-binding molecule in said first sample.