Fig. 3

Expression analysis of EYFP on transcript and enzyme level. A The EYFP reporter strains (Table 1) carrying the expression cassette at either the pyr4, the asl1, or the his1 locus were cultivated in a 12-well plate in 1.5 ml MAM containing glucose, lactose, glycerol, or xylan as carbon source. After incubation at 30 °C for 48 h, RNA was extracted, and cDNA was synthesized. The relative transcript levels of the eyfp were determined in a RT-qPCR assay using act1 and sar1 for normalization and the Pfaffl method [15] for calculation. QM6a eyfp(pyr4) on glucose was used as reference sample. The arithmetic average of all samples from all carbon sources are depicted in the bar chart. Error bars represent standard deviation. B The strains T. reesei QM6a Δpyr4, and the EYFP reporter strains carrying the expression cassette at either the pyr4, the asl1, or the his1 locus were cultivated in a fluorescence 96-well plate in MAM containing glucose, lactose, glycerol, or xylan as carbon sources. After incubation at 30 °C without agitation for 72 h, the total fluorescence (ex 490, em 510–570) was measured. Bars represent the arithmetic average of three independent replicates. Error bars represent standard deviation