RNA-Seq of RyhB R2
For RNA sequencing, cells were cultured in 1 L of medium to the exponential phase with an OD600 nm value of 0.8. After RNA was extracted, the Agilent RNA 6000 Nano Kit was used to assess the quality and quantity of RNA (Agilent, USA). Then mRNA was enriched by eliminating rRNA using the Ribo-ZeroTM Magnetic Kit (Epicentre). The ribosomal-depleted mRNA was fragmented and primed with random primers. Once the first-strand cDNA was formed, the second-strand cDNA was synthesized using DNA polymerase I, RNase H, dNTP, and buffer. Then, the QiaQuick PCR extraction kit (Qiagen, Germany) was used to purify cDNA fragments. After end-repairing and poly(A)-tailing, cDNA products were ligated to Illumina sequencing adapters, and fragments with a size of 300500 bp were selected.