Curved DNA fragments display retarded elution upon anion exchange HPLC.
Restriction fragments containing established curved regions, such as the pBR322 tn3 region, the phage lambda attP region and the SV40 T-antigen and terminus of replication regions, exhibit systematically retarded elution upon anion exchange based HPLC. Using this method, we were able to detect readily other SV40 curved regions, exhibiting also the polyacrylamide gel electrophoresis retardation anomaly, including several RNA polymerase initiation sites. Unlike gel retardation, HPLC retardation exhibited by curved DNA persists at 55 degrees C and is observed for fragments ranging from 150 bp up to 5 kb. The observed preferential attachment of DNA fragments containing curved DNA to the ionic groups of the column suggests a common dipole character of these regions due to the local accumulation of charges resulting apparently from the compression in the minor groove of curved DNA.
Fack
F
F
Institut de Biologie Moléculaire des Plantes du CNRS, Strasbourg, France.
Sarantoglou
V
V
D010582:Bacteriophage lambda / Q000235:genetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004247:DNA / Q000737:chemistry* / Q000302:isolation & purification; D004279:DNA, Viral / Q000737:chemistry* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000737:chemistry / Q000302:isolation & purification; D013539:Simian virus 40 / Q000235:genetics; D013696:Temperature
D004279:DNA, Viral; D009841:Oligonucleotides; D004247:DNA