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. Author manuscript; available in PMC: 2011 Sep 1.
Published in final edited form as: Mol Microbiol. 2010 Sep;77(6):1367–1379. doi: 10.1111/j.1365-2958.2010.07265.x

Figure 4. Effect of cas genes disruption, deletions and overproduction of processed CRISPR transcripts abundance.

Figure 4

A. A Northern blot showing abundance of spacer 4 transcript in E. coli strains with indicated cas genes “clean” deletions lacking the kanamycin-resistance gene present in original strains used in experiments shown in Figs. 1–3. RNA from one such strain, labeled ΔcasA_KanR, was loaded on the second lane. The probe was designed to reveal RNA produced by rightward (see Fig. 1A) transcription.

B. Changes in abundance of cas gene transcripts in cas disruption strains from the Keio collection were revealed by semi-quantitative RT-PCR as described in Experimental Procedures. In each vertical raw, the amounts of transcripts corresponding to genes indicated in the left are shown. The gyrB transcript was chosen as a control, whose level was not expected to depend on cas genes. Lane 10 shows results obtained with oligonucleotide pairs used for RT-PCR in lanes 1–9 using genomic DNA as a template (the reverse transcription step was omitted in this case)

C. A Northern blot showing abundance of spacer 4 transcripts in E. coli strains overexpressing indicated cas genes from ASKA library plasmids.