The purification of nuclease-free T4-RNA ligase.
RNA ligase has been highly purified in good yields from bacteriophage T4-infected Escherichia coli by a rapid and reproducible procedure. The enzyme is free of phosphomonoesterase and ribonuclease activities and is therefore suitable for the synthesis of oligoribonucleotides and for the labeling of the 3'-terminus of RNA. Greater than 90% of the protein in the enzyme preparation migrates as a single band on gradient polyacrylamide gels containing sodium dodecyl sulfate during electrophoresis. For use as a DNA synthesis reagent the enzyme may be reliably freed of deoxyribonuclease activity by an additional chromatographic procedure using a commercially avialable resin.
D002845:Chromatography; D003090:Coliphages / Q000201:enzymology*; D003851:Deoxyribonucleases / Q000378:metabolism*; D004926:Escherichia coli; D008970:Molecular Weight; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D011118:Polynucleotide Ligases / Q000302:isolation & purification*; D011130:RNA Ligase (ATP) / Q000302:isolation & purification*; D012260:Ribonucleases / Q000378:metabolism
D003851:Deoxyribonucleases; D012260:Ribonucleases; D010744:Phosphoric Monoester Hydrolases; D011118:Polynucleotide Ligases; D011130:RNA Ligase (ATP)