An Escherichia coli system for evolving improved light-controlled DNA-binding proteins.

Autor:	Mazumder M; Department of Chemical and Physical Sciences, University of Toronto Mississauga, 3359 Mississauga Rd. N., Mississauga, Ontario, Canada L5L 1C6., Brechun KE; Department of Chemistry, University of Toronto, 80 Saint George St, Toronto M5S 3H6, Canada., Kim YB; Department of Chemistry, University of Toronto, 80 Saint George St, Toronto M5S 3H6, Canada., Hoffmann SA; Institute for Biochemistry and Biology, University of Potsdam, Karl-Liebknecht-Str. 24-25, Potsdam, Golm 14476, Germany., Chen YY; Department of Chemistry, University of Toronto, 80 Saint George St, Toronto M5S 3H6, Canada., Keiski CL; Department of Chemistry, University of Toronto, 80 Saint George St, Toronto M5S 3H6, Canada., Arndt KM; Institute for Biochemistry and Biology, University of Potsdam, Karl-Liebknecht-Str. 24-25, Potsdam, Golm 14476, Germany., McMillen DR; Department of Chemical and Physical Sciences, University of Toronto Mississauga, 3359 Mississauga Rd. N., Mississauga, Ontario, Canada L5L 1C6 david.mcmillen@utoronto.ca awoolley@chem.utoronto.ca., Woolley GA; Department of Chemistry, University of Toronto, 80 Saint George St, Toronto M5S 3H6, Canada david.mcmillen@utoronto.ca awoolley@chem.utoronto.ca.
Jazyk:	angličtina
Zdroj:	Protein engineering, design & selection : PEDS [Protein Eng Des Sel] 2015 Sep; Vol. 28 (9), pp. 293-302. Date of Electronic Publication: 2015 Aug 05.
DOI:	10.1093/protein/gzv033
Abstrakt:	Light-switchable proteins offer numerous opportunities as tools for manipulating biological systems with exceptional degrees of spatiotemporal control. Most designed light-switchable proteins currently in use have not been optimised using the randomisation and selection/screening approaches that are widely used in other areas of protein engineering. Here we report an approach for screening light-switchable DNA-binding proteins that relies on light-dependent repression of the transcription of a fluorescent reporter. We demonstrate that the method can be used to recover a known light-switchable DNA-binding protein from a random library. (© The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.)
Databáze:	MEDLINE
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