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Vlatakis G. Skarpelis G. Stratidaki I. Bouriotis V. Clonis Y. D. 《Applied biochemistry and biotechnology》1987,15(3):201-212
The resolution of restriction endonucleases from the same microorganism is conventionally achieved by lengthy fractionation
protocols. We now report effective single-step procedures that exploit dye-ligand chromatography for the resolution and purification
of restriction enzymes. After suitable initial screening, we demonstrated that resolution of two restriction activites can
be achieved in one chromatographic step, and further purification can subsequently be effected using selected dye-adsorbents.
Accordingly, we resolved in one step, Hpa I from Hpa II, Hind II from Hind III, and Sac I from Sac II. Furthermore, a three-step
Chromatographic procedure has been developed to purify EcoRV suitable for commercial exploitation, as judged by the “overdigestion”
and “cut-ligate-recut” quality control tests. 相似文献
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