Fig. 4: BGCs deletions in S. coelicolor M1146. | Nature Communications

Fig. 4: BGCs deletions in S. coelicolor M1146.

From: Engineered CRISPR-Cas9 for Streptomyces sp. genome editing to improve specialized metabolite production

Fig. 4

a Representation of the plasmids constructed for BGC deletions in S. coelicolor using Cas9-BD. Insertion of sgRNA to create a cleavage of BGC, and two homologous regions for recombination of ~1 kb were inserted after cleavage into a plasmid containing the gene Cas9-BD. To perform multiple deletion, sets of sgRNA and homologous regions were inserted in the plasmids. b–d Scheme of BGC deletion of desferrioxamine B, SapB, and SCO-2138 and confirmation via nested PCR. e Result of double deletion of desferrioxamine B and SapB. f Triple deletion, including desferrioxamine B, SapB, and SCO-2138 BGC. All conjugations were performed in a single replicate (n = 1). Source data are provided as a Source Data file. Ctl, product of PCR amplification using the genomic DNA of S. coelicolor M1146 as the template. Mar, DNA ladder marker; Des. B, desferrioxamine B.

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