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

Authors: Cristina Hernandez Rollan, Kristoffer Bach Falkenberg, Maja Rennig, Andreas Birk Bertelsen, Morten Norholm

Summary: This protocol describes the expression and extraction of heterologously cytoplasmically expressed proteins in the bacterium E.coli, for T7 based expression systems. Specifically, it describes how to produce "hard-to-produce", as for example, LPMOs.It is based on a modified version of:"IPTG Induction and  Extraction of Proteins from Bacteria" by Swathi Arur and Sudhir Nayak, Schedl Lab. Washington University Genetics, St. Louis. Hemsworth, G. R., Henrissat, B., Davies, G. J., and Walton, P. H. (2014) Discovery and characterization of a new family of lytic polysaccharide monooxygenases. Nat. Chem. Biol.10, 122–126.The extraction of cytoplasmatic proteins is followed by a chemical lysis protocol, alternatively, sonication can also be employed.

Affiliations: Technical University of Denmark, Technical University of Denmark, Technical University of Denmark, Technical University of Denmark, Technical University of Denmark

Version: 1

Publication Date: 2020

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