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.
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Chhun, A., Alberti, F. (2022). CRISPR/Cas9-Based Methods for Inactivating Actinobacterial Biosynthetic Genes and Elucidating Function. In: Skellam, E. (eds) Engineering Natural Product Biosynthesis. Methods in Molecular Biology, vol 2489. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2273-5_11
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DOI: https://doi.org/10.1007/978-1-0716-2273-5_11
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