European Journal of Biochemistry · 1973 · 860 citations · 25 references
Life SciencesBiochemistryMolecular BiologyEnzyme TechnologyEnzymologyNucleic Acid BiochemistryEnzyme CharacterizationEnzyme IsolationEnzyme KineticsEnzyme PurificationFungal BiochemistryNucleic Acid PurificationAmylase BiologyEnzymatic Nucleic Acid ProcessingFungal EnzymologyMolecular Weight DeterminationNucleasesPh DependencePh Optima
The nuclease is useful for measuring annealing of labeled DNA. A simple purification scheme was devised to isolate Aspergillus nuclease S1 from commercial crude amylase powder. Purification yielded 90 % pure enzyme with a 27 % overall yield, and the metalloprotein (32 kDa) is strictly single‑strand‑specific, degrades both DNA and RNA, has a sharp pH optimum at 4.2, and remains active in low dodecylsulfate concentrations.
A simple scheme has been devised for the purification of Aspergillus nuclease S 1 from commercially available crude amylase powder. Five purification steps yield 90% pure nuclease with an overall yield of 27%. The enzyme is shown to be absolutely specific for single‐stranded nucleic acids. It degrades both DNA and RNA. The nuclease exhibits a sharp pH optimum at pH 4.2 and is active in low concentrations of dodecylsulfate. Aspergillus nuclease S 1 is a metalloprotein of molecular weight 32000. The utility of this nuclease for measuring annealing of labeled DNA is discussed.
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Nucleic acid reassociation in formamide
Betty L. McConaughy, Charles D. Laird, Brian J. McCarthy · Biochemistry · 1969 · 603 citations