Stimulation of intermolecular ligation with E. coli DNA ligase by high concentrations of monovalent cations in polyethylene glycol solutions.
In the presence of high concentrations of the nonspecific polymer polyethylene glycol (PEG), intermolecular cohesive-end ligation with the DNA ligase from Escherichia coli was stimulated by high salt concentrations: 200 mM NaCl or 300 mM KCl in 10% (w/v) PEG 6000 solutions, and 100-200 mM NaCl or 150-300 mM KCl in 15% PEG 6000 solutions. Intermolecular blunt-end ligation with this ligase was also stimulated at 100-150 mM NaCl or 150-250 mM KCl in 15% PEG 6000 solutions. The extent of such intermolecular ligation increased and the salt concentrations at which ligation was stimulated extended to lower concentrations when we raised the temperature from 10 to 37 degrees C.
D000249:Adenosine Monophosphate / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism*; D011088:DNA Ligases / Q000378:metabolism*; D004269:DNA, Bacterial / Q000378:metabolism*; D004270:DNA, Circular / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D008970:Molecular Weight; D010957:Plasmids; D011092:Polyethylene Glycols / Q000494:pharmacology; D011118:Polynucleotide Ligases / Q000378:metabolism*; D011188:Potassium / Q000494:pharmacology; D012964:Sodium / Q000494:pharmacology
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004270:DNA, Circular; D011092:Polyethylene Glycols; D000249:Adenosine Monophosphate; D012964:Sodium; D011088:DNA Ligases; D011118:Polynucleotide Ligases; D011188:Potassium