Transcriptomic analysis of the acuK and the acuM deletion mutants of Talaromyces marneffei

Identification of global target genes that their expression levels are dependent on the AcuK and the AcuM transcription factors from T. marneffei. Results provided information that the AcuK and AcuM are global regulators important for adaptation and defense against harsh environmental conditions, such as the response to iron limitation, utilization of alternative carbon sources, and antioxidation. Conidia of T. marneffei ATCC18224, ∆acuK and ∆acuM strains were inoculated at a final concentration of 108 cell/mL in 250 mL synthetic dextrose (SD) broth (1%glucose , 7 uM ferrous sulfate) and incubated at 25°C for 36 hours for the generation of mycelium mass. The cultures were harvested by centrifugation at 4500 rpm for 10 min. Approximately 2 g wet weight was homogenized in a TRIzol reagent (Invitrogen, Life Technologies, Carlsbad, CA, USA) by using a bead beater (Biospec, Bartlesville, OK, USA). Total RNA was isolated following the manufacturer’s protocol. The RNA was digested with 1 U/µL DNase I to eliminate the residual DNA and purified by isopropanol precipitation. RNA concentrations were determined with a spectrophotometer (Nanodrop 2000: Thermo Scientific, Waltham, MA, USA). RNA integrity and purity were tested by agarose gel electrophoresis. Library preparation and RNA sequencing were performed by a service from Novogene Co., Ltd. (Hong Kong).