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. 2019 Jun 7;20(11):2789. doi: 10.3390/ijms20112789

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Optimized schemes for expression, purification, and ATPase-assay of hexa-histidine (His)-tagged KaiC. (a) Overview of the protocol. (b) Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of His-tagged KaiC during its purification. Lane 1, molecular mass makers; lane 2, the pre-purified glutathione S-transferase (GST)-tagged KaiC; lane 3, the supernatant fraction after sonication; lane 4, the pellet fraction after sonication; lane 5, the proteins enriched on Ni-Separose affinity column and then eluted; lane 6; the main fraction eluted from Resource Q column; lane 7, the main fraction separated by Superdex 200 column. (c) Pseudo-parallel treatment of three sets of two KaiC mutants to achieve six KaiC mutants per week.