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. 2020 Mar 4;14(6):1479–1493. doi: 10.1038/s41396-020-0623-5

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — a The genetic organization of cas operon in subtype I-C CRISPR-Cas system loci from B. cereus VD115. b Plasmids derivatives of pHT-304 containing synthetic VD115 spacer 1 accompanied by 5′-TTN or 5′-NNN PAMs (TGC, TAC, GTC and ATC) were transformed into B. cereus VD115 strain and the transformation efficiency of plasmids was analyzed and showed as the number of CFU/mL. The immunity assays of subtype I-C CRISPR-Cas locus in B. cereus VD115 (c) and in B. thuringiensis BMB1685 (d) strains. The immunity was assessed via transformation of plasmids containing spacers flanked by a 5′-TTC PAM site in both strains. The transformation was conducted via electroporation, and the results were expressed as the number of CFU/mL. The pHT-304 vector was used as negative control in all assays. e T-A cloning and sequencing demonstrated the deletion region of cry6Aa gene. A loss of 198 bps in the edited cry6A (∆cry6Aa) is shown, compared with the wild type cry6Aa gene in BMB1685 strain. f The Cry6Aa protein production in BMB1685 strain and in selected cry6Aa deletion mutants was analyzed by SDS-PAGE. The red arrow points to the band of the complete Cry6Aa protein.