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. 2015 Feb 4;5:8248. doi: 10.1038/srep08248

Figure 2. Generation and identification of Fusarium graminearum myosin-5 gene recombinant mutants by gene replacement.

Figure 2

(a) Schematic representation of gene replacement strategy. The upper part represents the genomic locus target of the replacement construct. The black dashed lines represent the surrounding genomic region. The gene replacement cassette contains the hygromycin resistance gene and the myosin-5 mutated zone. Primer binding sites are indicated by arrows (see Table S1 for the primer sequences). The asterisk represents the selected myosin-5 mutated zone. (b) and (c) Polymerase chain reaction (PCR) strategy to screen replacement transformants: (b) PCR performed with primer pair A5/A6; a 1.9-kb amplified fragment indicates replacement integration at the left junction. (c) PCR performed with primer pair A7/A8; a 3.7-kb amplified fragment indicates replacement integration at the right junction. (d) Southern blot hybridization analysis of 2021 and all the mutants using a 622-bp myosin-5 upstream fragment as probe, and genomic DNA digested with SacI.