A rapid procedure to purify Escherichia coli DNA topoisomerase I |
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Authors: | Xu Xiaozhou Leng Fenfei |
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Institution: | aDepartment of Chemistry and Biochemistry, Florida International University, 11200 SW 8th Street, Miami, FL 33199, USA |
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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. |
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Keywords: | Escherichia coli DNA topoisomerase I DNA relaxation Q-Sepharose FF column SP-Sepharose FF column |
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