Simultaneous purification of DNA topoisomerase I and II from eukaryotic cells |
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Authors: | U Strausfeld A Richter |
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Institution: | Universit?t Konstanz, Fakult?t für Biologie, FRG. |
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Abstract: | We have developed a procedure for the simultaneous purification of DNA topoisomerase I and II from calf thymus. Both enzymes were first extracted from isolated nucleoprotein complexes. After batchwise chromatography on hydroxylapatite the two enzyme activities were separated on a FPLC phenylsuperose column. The enzymes were further purified by a second chromatography on phenylsepharose (topo I) or FPLC Mono Q (topo II). The purification can be finished within three days, yielding 0.5-1.0 mg quantities of homogeneous, enzymatic active preparations of the two proteins from 200 g of starting material. |
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