A rapid purification method of restriction endonucleases from Haemophilus strains.
A simple and rapid method of purification of restriction endonucleases from different Haemophilus strains is presented. By this method highly purified and stable enzymes can be obtained. Separation of different restriction activities present in the same strain is possible. This method was so far successfully used with Haemophilus influenzae, Haemophilus parainfluenzae and Haemophilus aegyptius strains. The main advantages over previously published procedures reside in the simplication of certain purification steps (for instance the BioGel A 0.5 M filtration is replaced by a hydroxyapatite batch step), elimination of exonuclease activity by fractionation with (NH4) 2SO4, separation of different restriction activities by phosphocellulose chromatography, application of this method to various strains and high purification degree of enzymes.
D002845:Chromatography; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004262:DNA Restriction Enzymes / Q000302:isolation & purification*; D004279:DNA, Viral; D004589:Electrophoresis, Disc; D004720:Endonucleases / Q000302:isolation & purification*; D006190:Haemophilus / Q000201:enzymology*; D006193:Haemophilus influenzae / Q000201:enzymology; D006882:Hydroxyapatites; D008722:Methods; D013539:Simian virus 40; D013045:Species Specificity
D004279:DNA, Viral; D006882:Hydroxyapatites; D004720:Endonucleases; D004262:DNA Restriction Enzymes