Specific protection of nucleotides in the lac operator from DMS methylation and DNase I nicking by crude bacterial cell extracts.
Crude bacterial cell extracts prepared from an Escherichia coli lacIq strain were shown to protect specific nucleotides in the lac operator from methylation by dimethyl sulfate (DMS) or digestion by DNase I, whereas no protection was observed using extracts prepared from a nearly isogenic lacI- strain. These experiments show that it is not necessary to use purified regulatory proteins in experiments designed to localize sequences on DNA which interact with proteins. Therefore, crude cell extracts should be useful in DNA "footprinting" experiments to define regions of DNA which bind to unknown regulatory proteins.
D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000235:genetics; D004268:DNA-Binding Proteins; D003850:Deoxyribonuclease I; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics*; D007763:Lac Operon; D008745:Methylation; D010957:Plasmids; D013463:Sulfuric Acid Esters / Q000378:metabolism*; D013464:Sulfuric Acids / Q000378:metabolism*
D002352:Carrier Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D013463:Sulfuric Acid Esters; D013464:Sulfuric Acids; D004706:Endodeoxyribonucleases; D004262:DNA Restriction Enzymes; D003850:Deoxyribonuclease I; C007482:dimethyl sulfate