Separation of G structures formed by a 27-mer guanosine-rich oligodeoxyribonucleotide by dye-ligand affinity chromatography.
G-DNA structures, formed by a 27-mer guanosine-rich oligodeoxyribonucleotide, AACCCGGCGTTCGGGGGGTACCGGGTT, were isolated and studied by dye-ligand affinity chromatography, using a Reactive Green 19-agarose resin (RG19-aga) and gel electrophoresis. The experiments were performed in the presence of Li+, Na+ and K+, which are able to stabilise the G structures to different extents. Desalting procedures followed by affinity chromatography, performed in the presence of Li+, gave us information on the relationships among the species isolated and their stability. The results show that the more stable species were those obtained in the presence of K+, while in the presence of Li+, the formation of G structures was negligible and the oligonucleotide was almost exclusively present as a stem-loop structure recognised by the RG19-aga affinity resin. Electrophoretic and denaturation and renaturation experiments supported the affinity chromatography results.
Alberghina
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Dipartimento di Scienze Chimiche, Università di Catania, Italy.
Fisichella
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Renda
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D001483:Base Sequence; D002846:Chromatography, Affinity / Q000379:methods*; D004396:Coloring Agents; D004587:Electrophoresis, Agar Gel; D006151:Guanosine / Q000032:analysis*; D008024:Ligands; D008094:Lithium / Q000494:pharmacology; D015394:Molecular Structure; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000302:isolation & purification*; D011188:Potassium / Q000494:pharmacology; D012685:Sepharose; D012964:Sodium / Q000494:pharmacology
D004396:Coloring Agents; D008024:Ligands; D009838:Oligodeoxyribonucleotides; D006151:Guanosine; D012685:Sepharose; D008094:Lithium; D012964:Sodium; D011188:Potassium