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Fig. 3 | Hereditas

Fig. 3

From: Transformation and gene-disruption in the apple-pathogen, Neonectria ditissima

Fig. 3

A The GFP expression vector, pCT74, developed by Lorang et al., (2001) was used during the transformation. The primers T7 and T3 were used to amplify 3117 bp containing the coding sequence for the GFP expression and hygromycin selection, used during the transformation. B Disruption construct generated by GeneArt (Thermo Scientific). Primer Set 1 (Table 2) was used to amplify the coding sequence (3548 bp) from the NRPS gene, AK830_g4721 and cloned into pJet1.2. The backbone of the plasmid with a 5′-fragment (1100 bp) and a 3′-fragment (1200 bp) from the coding sequence at each end, was amplified with Primer Set 3 (Table 2), while Primer Set 4 (Table 2) was used to amplify a linear fragment containing the hygromycin cassette from plasmid pJCA-HygII (Vélëz et al., unpublished). The fragments were ligated using the GeneArt® Seamless Cloning and Assembly Enzyme Mix (Invitrogen). Figure generated using SnapGene® software (Insightful Science; snapgene.com)

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