The Δpmk1 and Δmofim1 deletion mutants were generated according to previous methods (Liu et al. 2022 (link)). Briefly, two approximately 1.5 kb fragments of the sequences flanking PMK1 were amplified with two primer pairs. The obtained DNA fragments were ligated upstream and downstream of the hygromycin gene. Then, the recombinant DNA was infused into the pGKO vector (Zhou et al. 2017 (link)) via homologous recombination cloning (ClonExpress MultiS One Step Cloning Kit, Vazyme Biotech, C112). The protoplasts of wild-type Y34 were transformed with pGKO-PMK1 for targeted gene deletion.
To construct plasmids expressing PMK1-mCherry and MoFim1-GFP, approximately 1.5 kb of the native promoter region from the M. oryzae genome was amplified and cloned and inserted into the pKNTG expression vector (Zheng et al. 2016 (link)). All the constructs were cloned by homologous recombination (ClonExpress MultiS One Step Cloning Kit, Vazyme Biotech, Nanjing, China; C112); all the primers with restriction enzyme sites are listed in Supplemental Table S1 .
To construct plasmids expressing PMK1-mCherry and MoFim1-GFP, approximately 1.5 kb of the native promoter region from the M. oryzae genome was amplified and cloned and inserted into the pKNTG expression vector (Zheng et al. 2016 (link)). All the constructs were cloned by homologous recombination (ClonExpress MultiS One Step Cloning Kit, Vazyme Biotech, Nanjing, China; C112); all the primers with restriction enzyme sites are listed in Supplemental Table S
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