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.

The Adaptive Laboratory Evolution (ALE) experiment allowed to select Corynebacterium glutamicum strain GluA T5, which grew faster and produced more glutarate than the parent strain GluA T0, and strain GluA T7 that grew as fast as GluA T5, but produced more 5AVA than GluA T5. To explore differences in the gene expression in the evolved strains, C. glutamicum GluA T0, GluA T5 and GluA T7 biological triplicates 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. RNA isolation, purification and quality control was performed as described [82] and the high quality RNA (RNA integrity number > 9.0) was kept at -80°C until further use. Ribo-Zero rRNA Removal Kit (Bacteria) from Illumina (San Diego, CA, USA) was used to remove the ribosomal RNA molecules from the isolated total RNA. Preparation of cDNA libraries were performed according to the manufacturer’s instructions of TruSeq stranded mRNA Kit (Illumina, San Diego, USA). Subsequently, each cDNA library was sequenced on a HiSeq1500 (2 x 70nt PE rapid v2) and NextSeq 500 (2 x 75nt PE mid output v2.5) Sequencer system (Illumina, San Diego, USA).