[Use of Cibacron blue F 3GA-CL-sepharose 6B for the purification of T4 DNA- and RNA-ligases].
The sorption capacity of the dye cibacron blue F3GA, immobilized on CL-Sepharose 6B and other support matrices, in respect to DNA- and RNA-ligases T4 was being studies. Cibacron blue F3GA immobilized on CL-Sepharose 6B binds a three-fold amount of DNA-ligase in comparison to RNA-ligase. The enzyme chromatography on cibbacron blue F3GA-CL-Sepharose 6B revealed a stronger linkage between DNA-ligase T4 and the sorbent than between RNA-ligase T4 and the sorbent. Elution was performed with potassium chloride. DNA-ligase T4 was eluted with 0.25-0.5 M KCl and RNA-ligase T4 with 0.08-0.18 M KCl. Since deoxyexonuclease contaminants possess stronger bonds with the sorbent than ligases, elution of deoxyexonucleases occurs at higher concentrations of KCl. Chromatography of enzymes on cibacron blue F3GA-CL-Sepharose 6B allows one to obtain DNA- and RNA-ligases essentially free of DNase and RNase contaminants.
D000042:Absorption; D002845:Chromatography / Q000379:methods; D011088:DNA Ligases / Q000032:analysis / Q000302:isolation & purification*; D006882:Hydroxyapatites / Q000494:pharmacology; D010744:Phosphoric Monoester Hydrolases / Q000032:analysis / Q000302:isolation & purification; D011118:Polynucleotide Ligases / Q000302:isolation & purification*; D011130:RNA Ligase (ATP) / Q000032:analysis / Q000302:isolation & purification*; D012260:Ribonucleases / Q000032:analysis / Q000302:isolation & purification; D012685:Sepharose / Q000031:analogs & derivatives* / Q000494:pharmacology; D014227:Triazines / Q000494:pharmacology*
D006882:Hydroxyapatites; D014227:Triazines; C012842:Cibacron Blue F 3GA; C047264:sepharose CL 6B; D012685:Sepharose; D012260:Ribonucleases; D010744:Phosphoric Monoester Hydrolases; D011088:DNA Ligases; D011118:Polynucleotide Ligases; D011130:RNA Ligase (ATP)
Ispol'zovanie CL-sefarozy 6B-tsibakron golubogo F3GA dlia ochistki DNK- i RNK-ligaz T4.