Simple isolation of DNA hydrophobically complexed with presumed gene regulatory proteins (M3).
Chromatin from chicken reticulocytes and mouse Ehrlich ascites tumor cells has been extracted with 2 M NaCl, leaving a portion of the DNA still complexed with a fraction of nonhistones (designated M3, since it can be dissociated from DNA in solutions of 3 M NaCl containing 5 M urea). The DNA complexed with M3, separated from the bulk DNA by centrifugation, was found to contain sequences poorly represented in bulk DNA. Specifically we found that DNA--M3 complexes isolated from chicken reticulocyte chromatin were enriched in globin gene sequences by 20-fold relative to unfractionated DNA and by over 1000-fold relative to DNA rendered free of protein following the extraction of chromatin with 2 M NaCl. We have therefore isolated DNA fractions complexed with M3 which are enriched in specific sequences as may be determined by M3.
D000818:Animals; D002286:Carcinoma, Ehrlich Tumor / Q000378:metabolism; D002645:Chickens; D002843:Chromatin / Q000032:analysis; D004247:DNA / Q000302:isolation & purification* / Q000378:metabolism; D004262:DNA Restriction Enzymes; D004273:DNA, Neoplasm / Q000302:isolation & purification* / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D005809:Genes, Regulator; D007700:Kinetics; D051379:Mice; D009693:Nucleic Acid Hybridization; D009698:Nucleoproteins / Q000302:isolation & purification*; D012156:Reticulocytes / Q000378:metabolism; D013698:Templates, Genetic
D002843:Chromatin; D004273:DNA, Neoplasm; D009698:Nucleoproteins; D004247:DNA; D012321:DNA-Directed RNA Polymerases; D004262:DNA Restriction Enzymes