Abstract
On the basis of the asymmetrical charge distribution of Escherichia coli DNA topoisomerase I, we developed a new procedure to purify E. coli DNA topoisomerase I in the milligram range. The new procedure includes using both cation- and anion-exchange columns, i.e., SP-Sepharose FF and Q-Sepharose FF columns. The E. coli DNA topoisomerase I purified here is free of DNase contamination. The kinetic constants of the DNA relaxation reaction of E. coli DNA topoisomerase I were also determined.
Copyright © 2011 Elsevier Inc. All rights reserved.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Bacterial Proteins / genetics
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Bacterial Proteins / isolation & purification
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Bacterial Proteins / metabolism*
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Chromatography, Ion Exchange
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Cloning, Molecular
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DNA / metabolism
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DNA Topoisomerases, Type I / genetics
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DNA Topoisomerases, Type I / isolation & purification
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DNA Topoisomerases, Type I / metabolism*
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Deoxyribonucleases / metabolism
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Electrophoresis, Polyacrylamide Gel
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Escherichia coli / enzymology*
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Escherichia coli / genetics
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Gene Expression
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Kinetics
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Plasmids
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism*
Substances
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Bacterial Proteins
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Recombinant Proteins
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DNA
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Deoxyribonucleases
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DNA Topoisomerases, Type I