The purification of nuclease-free T4-RNA ligase

Biochim Biophys Acta. 1979 Mar 28;562(1):149-61. doi: 10.1016/0005-2787(79)90134-5.

Abstract

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.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Chromatography
  • Coliphages / enzymology*
  • Deoxyribonucleases / metabolism*
  • Escherichia coli
  • Molecular Weight
  • Phosphoric Monoester Hydrolases / metabolism
  • Polynucleotide Ligases / isolation & purification*
  • RNA Ligase (ATP) / isolation & purification*
  • Ribonucleases / metabolism

Substances

  • Deoxyribonucleases
  • Ribonucleases
  • Phosphoric Monoester Hydrolases
  • Polynucleotide Ligases
  • RNA Ligase (ATP)