Aspergillus oryzae Transcriptome or Gene expression. Aspergillus oryzae

In order to evaluate the role of HacA in coping with ER stress and as possible regulator of theunfolded protein response inA.oryzae, we proceeded to characterize hacAknockout (HacA-DE) and hacA in constitutively activeexpression (HacA-CA) strains. To construction hacAknockout strain, we generated oneby replacing thehacA(AO090124000074)coding region with a pyrG cassette. Correct integration of thecassette and replacement of thehacA sequence was verified by PCR andthe absence of hacA was confirmed in theHacA-DE strain.To obtain astrain with a constitutively activated HacA transcription factor, we integrated the spliced form ofhacAthatlacks the 20 nucleotide intron intoHacA-DE strain hacA loci. Transformants withthe correct integration pattern wereselected after PCR analysis and the hacA gene in constitutively active was successfully expressed in theHacA-CA (ΔhacA:: hacA-i)strain.The isolated mRNAs of niaD300, HacA-DE and HacA-CAwere subjected to RNAsequencing.