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. 2018 Jul 20;10(7):301. doi: 10.3390/toxins10070301

Figure 2.

Figure 2

Figure 2

Strategy and confirmation of the mutant strains. (A) The scheme of rum1 deletion and complementation strategy (H: HindIII, probe: the probe used in southern blot analysis). (B) Gene knockout and complemented strains were verified by PCR analysis. (The rum1 ORF was confirmed by primers rum1-p9 and rum1-p10, AP fragment was confirmed by primers rum1-p1 and P801, and fragment BP was confirmed by primers P1020 and rum1-p4). (C) q-PCR verification of rum1 gene deletion and complementation strains. The actin gene was used as an inner reference. (D) Southern blot analysis for ∆rum1 mutant. Genomic DNA from above strains was digested with HindIII and hybridized with a 1.4 kb probe of the downstream region fragment of rum1 (3′-UTR) (The probe fragment was amplified with primers rum1-p3 and rum1-p4), (E) q-PCR analysis of the expression level of rum1 gene in ∆rum1 WT and ∆rum1-C strains. *** represents significant difference (p < 0.001).