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. 2013 Jun 14;2(4):595–609. doi: 10.1002/mbo3.94

Figure 5.

Figure 5

Cellulase gene expression and activity by misexpression of clr-2 in N. crassa. (A) K-means clusters of genes (Nc1-4) induced under Avicel or no carbon conditions in WT, under Avicel conditions in a Δclr-2 mutant and under no carbon media conditions for a strain with constitutive expression of clr-2 (Pc2). Heat map displays log (FPKM) scaled from min expression (bright blue) to the maximal expression (bright yellow). (B) Total expression of genes encoding major CAZy classes of Pc2 on sucrose versus no carbon media, as compared to WT on sucrose, no carbon media, or Avicel. Note the similarity in CAZy expression class in WT on Avicel versus the Pc2 strain on media lacking a carbon source. A Δclr-2 strain on Avicel shows a low induction of genes encoding some hemicellulases, due to slight contamination of Avicel with hemicellulose (Znameroski et al. 2012). (C) Coomasie-stained SDS PAGE gel of culture supernatants from WT,Δclr-2 (Δ), and Pc2 strains shifted to Avicel (4 days), sucrose (2 days), or media lacking a carbon source (2 days). Secreted proteins labeled from molecular weights and intensity patterns are derived from (Tian et al. 2009; Phillips et al. 2011ab2011b; Znameroski et al. 2012). Note similarity of secreted protein profile of the Pc2 strain grown on sucrose or no carbon as compared to Avicel-exposed WT and Pc2 cultures. (D) A time course of CMCase activity of supernatants from WT,Δclr-2, and Pc2 cultures after shift from sucrose pregrown cultures (16 h) to either Avicel or sucrose.