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URL: https://dx.doi.org/10.17504/protocols.io.k68czhw

Authors: Anika Wiegard

Group: Axmann Lab

Summary: This protocol can be used for:(i) heterologous expression of GST-tagged proteins from pGEX-6P1 based expression vectors in E. coli.(ii) purification of recombinant proteins via affinity chromatography using glutathione-agarose or glutathione-sepharose (GST tagged protein can be eluted with glutathione. Alternatively, the tag can be cleaved off by prescission protease)(iii) further purification of the eluted protein via anion exchange chromatographyThis protocol was modified fromWiegard A, Dörrich AK, Deinzer HT, Beck C, Wilde A, Holtzendorff J, Axmann IM: Biochemical analysis of three putative KaiC clock proteins from Synechocystis sp. PCC 6803 suggests their functional divergence. Microbiology 2013, 159, 948-958Snijder J, Schuller JM, Wiegard A, Lössel, P, Schmelling NM, Axmann IM, Plitzko JM, Förster F, Heck AJR: Structures of the cyanobacterial circadian oscillator frozen in a fully assembled state. Science 2017, 355(6330):1181-1184

Affiliations: Institute for Synthetic Microbiology, CEPLAS, Heinrich Heine University Duesseldorf, Germany

Version: 1

Publication Date: 2018

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