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. 2014 Sep 15;9(9):e107657. doi: 10.1371/journal.pone.0107657

Figure 2. The disruption of MrKu70 in M. robertsii using the herbicide-resistance gene Sur as the selection marker.

Figure 2

(A) The disruption plasmid of MrKu70 (bottom) and the relative position of MrKu70 in the wild-type strain. (B) Confirmation of the disruption of MrKu70 by PCR in the mutants with glufosinate ammonium resistance and without a GFP signal. 1 and 2 designate two different ΔMrKu70 mutants, and C is the wild-type strain. Top panel: PCR conducted with the primers Sur5 and MrKu70CF2, PCR products can be obtained only from MrKu70 disruption mutants; Bottom panel: PCR conducted using primers MrKu70CF1 and MrKu70CF2, and PCR products can be obtained in any strains other than MrKu70 disruption mutants. (C) Confirmation of the complementation of ΔMrKu70 by PCR using the primers MrKu70ORF-5 and MrKu70ORF-3 to amplify across the deleted region. 1 to 3: 3 different transformants with ΔMrKu70 complemented; C: the wild-type strain; M: ΔMrKu70; L: DNA ladder (DL 10004) from Generay (Shanghai, China).