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. 2011 Aug 3;39(21):9275–9282. doi: 10.1093/nar/gkr606

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© The Author(s) 2011. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — CRISPR3/Cas system of S. thermophilus provides immunity against plasmid transformation in E. coli cells. (A) Schematic representation of CRISPR3/Cas system cloning and construction of the plasmids for interference assay. Streptococcus thermophilus CRISPR3/Cas system was cloned into E. coli plasmid pACYC184. Plasmids for interference assays were obtained by inserting a proto-spacer and PAM into pUC18 plasmid. (B) Schematic representation of the plasmid transformation interference assay. Escherichia coli RR1 recipient strains carrying plasmids pCRISPR3 and pACYC184 with and without the S. thermophilus CRISPR3/Cas system, respectively, were transformed with plasmids pSP1 and pSP2 carrying proto-spacers and PAMs or pUC18. (C) Interference of plasmid transformation by S. thermophilus CRISPR3/Cas system in E. coli cells. Transformation efficiency is expressed as cfu per nanogram of plasmid DNA (mean ± SD).