characteristics.text: 2012-09-30T17:02:28Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2012-08-02T12:55:16Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA1523773
characteristics.characteristic: External Id
characteristics.text: Leibniz-Institute for Natural Product Research and Infection Biology Hans Knoll Institute
characteristics.characteristic: INSDC center name
characteristics.text: 2012-09-30T17:02:28Z
characteristics.characteristic: INSDC first public
characteristics.text: 2012-08-02T12:55:16Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS159692
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-1236:WT3
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: Protocols: Total RNA was isolated from wild-type and mpkA strains cultured in Aspergillus minimal medium (AMM) for 16 h at 37 C (Jain et al., 2011), using the Qiagen RNeasy Plant Mini kit (Qiagen, Germany), according to the manufacturers instruction. Transformation of A. fumigatus KU80D with plasmid pCR2.1DmpkA/Ura to generate the mpkA deletion mutant was carried out using protoplasts as previously described (Weidner et al., 1998). Transformation of A. fumigatus strain DmpkA for complementation of the deletion was per- formed as follows: conidia of the DmpkA strain were plated on AMM agar plates supplemented with 1 g/L 5-fluoroo- rotic acid (5-FOA, Fermentas, Germany) and 10 mM uracil (Fluka, Germany), in order to obtain DmpkA pyrGA strains. DmpkA pyrGA colonies were screened by Southern blot analysis for loss of the pyrG gene. Plasmid delta mpkA/Ura was used to transform DmpkA pyrGprotoplasts. Transformants were analyzed by PCR and Southern blot analysis for mpkA complementation. A posi- tive transformant strain was designated mpkAc. Plasmid pUCPyrG2mpkAp-lacZ, containing the mpkA promoter fused in frame with the E. coli lacZ gene, was used to transform A. fumigatus strain KHDpyrG (Sugareva et al., 2006). Southern blot analysis of the resulting trans- formant strain MpkAp-lacZ indicated the presence of the gene fusion in single copy at the pyrG gene locus. (Valiante V, Heinekamp T, Jain R, Haertl A, Brakhage AA: The mitogen-activated protein kinase MpkA of Aspergillus fumigatus regulates cell wall signaling and oxidative stress response. Fungal Genet Biol 2008, 45: 618-627.) Total RNA was isolated from wild-type and mpkA strains. Total RNA was used for Illumina next-generation sequencing (Bentley et al., 2008). For library preparation 5ug of total RNA per sample were processed using Illumina mRNA-Seq sample prep kit (RS-100-0801) following the manufacturers instruction
characteristics.characteristic: description
characteristics.text: transformation
characteristics.characteristic: geneticmodification
characteristics.text: wild_type
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/SO_0000817
characteristics.characteristic: genotype
characteristics.text: Aspergillus fumigatus
characteristics.characteristic: organism
characteristics.text: Aspergillus fumigatus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_746128
characteristics.characteristic: organism
characteristics.text: Aspergillus fumigatus
characteristics.characteristic: scientific_name
characteristics.text: WT3
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0001103
characteristics.characteristic: title