Illumina HiSeq 1500 paired end sequencing; RNAseq analysis of global gene expression patterns in Corynebacterium glutamicum Adaptive Laboratory Evolution (ALE) strains GluA T5 and GluA T7 with accelerated growth and glutarate production.
RNAseq analysis of global gene expression patterns in Corynebacterium glutamicum Adaptive Laboratory Evolution (ALE) strains GluA T5 and GluA T7 with accelerated growth and glutarate production.
C. glutamicum GluA T0, GluA T5 and GluA T7 were grown in CGXII minimal medium with 4% (w/v) glucose supplemented with 1 mM IPTG. Exponentially growing cells were harvested by centrifugation (14000 × g, 1 min) and kept at -80°C.  C. glutamicum GluA T0, GluA T5 and GluA T7 were grown in CGXII minimal medium with 4% (w/v) glucose supplemented with 1 mM IPTG. Exponentially growing cells were harvested by centrifugation (14000 × g, 1 min) and kept at -80°C. Strain reference: Pérez-García F, Jorge JMP, Dreyszas A, Risse JM, Wendisch VF. Efficient Production of the Dicarboxylic Acid Glutarate by Corynebacterium glutamicum via a Novel Synthetic Pathway. Front Microbiol 2018;9. https://doi.org/10.3389/fmicb.2018.02589. Total RNA was isolated from 3 biological replicates of C. glutamicum cells grown to the exponential phase using a Quick-RNA Miniprep Plus kit according to the manufacturer's instructions (Zymo Research). After additional DNase treatment, RNA samples were purified with an RNA Clean&Concentrator-5 kit (Zymo Research) and quantified with a DropSense 16 (Trinean). The quality of total RNA was controlled with an RNA 6000 Nano kit in an Agilent 2100 Bioanalyzer (Agilent Technologies).  To construct the whole transcriptome cDNA library, 2.5 μg total RNA (RIN > 9) was used for the depletion of rRNA with a Ribo-Zero rRNA Removal Kit (Bacteria) according to manufacturer's instructions (Illumina). The rRNA removal was checked with an Agilent RNA Pico 6000 kit and the Agilent 2100 Bioanalyzer (Agilent Technologies). The mRNA obtained was converted to a cDNA library according to the TruSeq Stranded mRNA Sample Preparation guide (Illumina). The quality and quantity of the cDNA library was checked with an Agilent High Sensitivity DNA kit and the Agilent 2100 Bioanalyzer (Agilent Technologies).