Fig. 3.

Fig. 3. Capturing the fungal gene cluster with CRISPR/Cas9 cleavage system. (A) SgRNA pairs designed to target Nrc gene cluster from N. fischeri. (B) Verification of the obtained construct pYJF31 containing Nrc gene cluster by restriction endonuclease digestion. Lane M: marker; Lanes 1-4: plasmids digested by KpnI. Lengths of target fragments are 15,631 bp, 6,209 bp, 4,432 bp, 2,782 bp and 2,388 bp; Lane 5-8: plasmids digested by NcoI. Lengths of target fragments are 7,600 bp, 7,348 bp, 6,271 bp, 5,975 bp, 2,511 bp and 1,737 bp. (C) SgRNA pairs designed to target genome Pfma gene cluster from P. fici. (D) Genomic DNA of P. fici analyzed using PFGE. Lane M: marker; Lane 1: P. fici gDNA without Cas9 cleavage; Lane 2: P. fici gDNA cleaved by Cas9 in low melting agarose plugs.

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