An Escherichia coli system for evolving improved light-controlled DNA-binding proteins
- 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.
| Author details: | Mostafizur Mazumder, Katherine E. BrechunORCiD, Yongjoo B. Kim, Stefan A. Hoffmann, Yih Yang Chen, Carrie-Lynn Keiski, Katja Maren ArndtORCiDGND, David R. McMillen, G. Andrew Woolley |
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| DOI: | https://doi.org/10.1093/protein/gzv033 |
| ISSN: | 1741-0126 |
| ISSN: | 1741-0134 |
| Pubmed ID: | https://pubmed.ncbi.nlm.nih.gov/26245690 |
| Title of parent work (English): | Protein engineering design & selection |
| Publisher: | Oxford Univ. Press |
| Place of publishing: | Oxford |
| Publication type: | Article |
| Language: | English |
| Year of first publication: | 2015 |
| Publication year: | 2015 |
| Release date: | 2017/03/27 |
| Tag: | directed evolution; fluorescent reporter; optogenetics |
| Volume: | 28 |
| Issue: | 9 |
| Number of pages: | 10 |
| First page: | 293 |
| Last Page: | 302 |
| Funding institution: | Natural Sciences and Engineering Research Council of Canada; Human Frontiers Science Program |
| Organizational units: | Mathematisch-Naturwissenschaftliche Fakultät / Institut für Biochemie und Biologie |
| Peer review: | Referiert |
