CRISPR/Cas9-Based Methods for Inactivating Actinobacterial Biosynthetic Genes and Elucidating Function
Details
Serval ID
serval:BIB_FC333623D005
Type
A part of a book
Publication sub-type
Chapter: chapter ou part
Collection
Publications
Institution
Title
CRISPR/Cas9-Based Methods for Inactivating Actinobacterial Biosynthetic Genes and Elucidating Function
Title of the book
Engineering Natural Product Biosynthesis
Publisher
Springer US
ISBN
9781071622728
9781071622735
9781071622735
Publication state
Published
Issued date
2022
Peer-reviewed
Oui
Volume
2489
Series
Methods in Molecular Biology
Pages
201-222
Language
english
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.
Keywords
Actinobacteria/genetics, CRISPR-Cas Systems/genetics, Gene Editing/methods, Streptomyces/genetics, Streptomyces/metabolism, Actinobacteria, Biosynthetic gene, CRISPR/Cas9, Natural products, pCRISPomyces-2
Pubmed
Create date
17/05/2022 13:21
Last modification date
12/04/2024 8:45
Usage data
