Fig. 2

Construction of transformants and effects of the truncated type ACE3-723 versus the native type ACE3-734 on cellulase production of T. reesei Rut-C30. a Schematic representation of A723 and A734 transformants. LML 2.1 is the erasable hygromycin selection marker in T. reesei. A734 transformants carry the native ACE3-734 as the test strains. A723 transformants bear ACE3-723 as controls. The black square denotes the loxP site left at the C-terminus of ACE3 after the marker was excised. The primers ace3-CF and D70-4 and HG3.6 and ace3-CR were used to verify the genotype of ACE3. b, c Effects of the truncated type ACE3-723 versus the native type ACE3-734 on cellulase production of T. reesei. The pNPCase activity of T. reesei Rut-C30 and transformants was examined after culture in liquid 2× Mandels’ medium with 2% lactose (b) or 1% Avicel (c). d–f Transcription of genes encoding the major cellulase (cbh1) and essential transcription factors for cellulase (ace3 and xyr1) was evaluated. Three independent experiments with three biological replicates each were performed. The sar1 gene was used as the internal control for normalization. Values are the mean ± SD of the results from three independent experiments. Asterisks indicate a significant difference (*p < 0.05, Student’s t test)