Comparison and Validation of Putative Pathogenicity-Related Genes Identified by T-DNA Insertional Mutagenesis and Microarray Expression Profiling in Magnaporthe oryzae

Figure 1

Southern blot and RT-PCR analysis of the transformants. (a) MGG_00623. DNA samples were digested with ApaI, probed with a 1.2 kb upstream flanking sequence fragment of the gene replacement vector. The 5.6 kb band was detected in mutant, whereas the 1.6 kb band was detected in wild-type Guy11; (b) MGG_00871. DNA samples were digested with KpnI, probed with a 1.0 kb upstream flanking sequence fragment of the gene replacement vector. The 7.4 kb band was detected in mutant, whereas the 3.8 kb band was detected in wild-type Guy11; (c) MGG_06704. DNA samples were digested with SacI, probed with a 1.3 kb downstream flanking sequence fragment of the gene replacement vector. The 11.4 kb band was detected in mutant, whereas the 9.3 kb band was detected in wild-type Guy11; (d) MGG_00745. DNA samples were digested with EcoRV, probed with a 1.0 kb upstream flanking sequence fragment of the gene replacement vector. The 3.5 kb band was detected in mutant, whereas the 2.5 kb band was detected in wild-type Guy11; (a–d) Lane of 1: Guy11; 2: transformant with selection marker but not single copy; 3: mutant with targeted gene had been replaced. (e) Lane of A: ΔMGG_00623; B: ΔMGG_00871; C: ΔMGG_06704; D: ΔMGG_00745.

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