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CRISPR/Cas9-based methods for inactivating actinobacterial biosynthetic genes and elucidating function
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Chhun, Audam and Alberti, Fabrizio (2022) CRISPR/Cas9-based methods for inactivating actinobacterial biosynthetic genes and elucidating function. In: Skellam, Elizabeth, (ed.) Engineering Natural Product Biosynthesis. Methods in Molecular Biology, 2489 . New York: Humana Press, pp. 201-222. ISBN 9781071622728
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Official URL: http://dx.doi.org/10.1007/978-1-0716-2273-5_11
Abstract
The CRISPR/Cas9 technology allows fast and marker-less genome engineering that can be employed to study secondary metabolism in actinobacteria. Here, we report a standard experimental protocol for the deletion of a biosynthetic gene in a Streptomyces species, using the vector pCRISPomyces-2 developed by Huimin Zhao and collaborators. We also describe how carrying out metabolite analysis can reveal the putative biosynthetic function of the inactivated gene.
| Item Type: | Book Item | ||||
|---|---|---|---|---|---|
| Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||
| Series Name: | Methods in Molecular Biology | ||||
| Publisher: | Humana Press | ||||
| Place of Publication: | New York | ||||
| ISBN: | 9781071622728 | ||||
| ISSN: | 1064-3745 | ||||
| Book Title: | Engineering Natural Product Biosynthesis | ||||
| Editor: | Skellam, Elizabeth | ||||
| Official Date: | 7 May 2022 | ||||
| Dates: |
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| Volume: | 2489 | ||||
| Page Range: | pp. 201-222 | ||||
| DOI: | 10.1007/978-1-0716-2273-5_11 | ||||
| Status: | Peer Reviewed | ||||
| Publication Status: | Published | ||||
| Access rights to Published version: | Restricted or Subscription Access |
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