Fig. 2

Properties of the hax1 locus and its encoded regulatory factor. a Locus targeted by amdS integration in QM9414_Dhax1 strains and its genetic environment according to JGI genome database of Trichoderma reesei QM6a v2.0. The targeted locus hax1, the downstream coding region referred to as “hypothetical protein” (hyp. protein) and the upstream gene encoding 2-isopropylmalate synthase are illustrated as yellow, light and dark blue arrows, respectively. b Schematic illustration of the 782 bp hax1 structural gene (black arrow) predicted by the web server AUGUSTUS. Start codon (ATG, red arrow), introns (grey boxes), site targeted by integration of the amdS cassette (yellow triangle), 5′-region rich in Xyr1-binding sites (green box) and position of the primers used for transcript analysis (thin, black arrows) are indicated. Numbers on top indicate position from ATG in base pairs. c Transcript levels of hax1 in QM9414 that was pre-grown and transferred to medium without carbon source (violet bars) or 1% (w/v) d-glucose (red bars) for 3 h. Indicated primer pairs were used for PCR 1, 2, 3, 4, and 5, respectively. Transcript levels were normalized to act and sar1, refer to PCR 1 (no carbon source) and are given in logarithmic scale (lg). Analysis was performed in technical triplicates. Error bars indicate standard deviations. Up_1, up-hax1 for_1; up-int, hax1 rev up-Intron; up_2, up-hax1 for_2; r1.int, hax1 rev 1.Intron; f1.int, hax1 for_1.Intron; rev-k, hax1 rev kurz; i-int, hax1 for_inter-Intron; d-int, hax1 rev_down-intron; for 3, hax1 for_3; rev, hax1 rev; n.d., not detectable. d In silico structure prediction of the lncRNA HAX1 with minimized free energy. Structure stability is increasing from violet, blue, green and yellow to red