Transcriptional Profiling of Metabolic Genes

The transcriptional expression of maeB, ppsA, ppc, pckA, pgi, zwf, gltA, and arcA in each strain was quantified using real-time PCR. Total RNA was isolated from individual cultures using a NucleoSpin RNA column (Takara Bio, Shiga, Japan) according to the manufacturer's protocol. Reverse transcription reactions and quantitative real-time PCR was performed using an Mx3005P Real-Time QPCR System (Agilent Technologies, Santa Clara, CA, USA) with RNA-direct SYBR Green Realtime PCR Master Mix (TOYOBO, Osaka, Japan). The primer pairs are listed in Supplementary Data 7. The normalized transcriptional level of each mRNA was calculated by the relative quantification method using the mdoG gene (coding glucan biosynthesis protein G) as the housekeeping gene^{61 (link)}.