Reporter assays for transcriptional regulation by YdeO.
[A] YdeO-expression induces the expression of target promoters. YY0201/pLUXnhaR (nhaR-lux), HYAA-JL (hyaA-lacZ), YY1101 (yiiS-lacZ), YY0201/pLUXgadWp (gadW-lux), YY0201/pLUXgadE (gadE-lux), and YY0201/pLUXslpp (slp-lux), and were transformed with pTrc99A (vector, white bar) and pYdeO (ydeO, black bar). Transformants grew until logarithmic phase and β-galactosidase and luciferase activities of cultures were measured as described in Materials and methods. The data show the average of independent eight experiments with standard deviation as the ratio of a vector-transformant. [B] APPC-JL (appC-lacZ) was transformed with pTrc99A (vector) or pYdeO (ydeO). Transformants grew until logarithmic phase and β-galactosidase was measured as decribed in [A]. The data show the average of independent eight experiments with standard deviation as the Miller unit. [C] The ydeO expression induced under anaerobic conditions. The activity of ydeO promoter was measured in YY0101 growing in LB medium with pH 7.2 and 5.5 under aerobic (+) and anaerobic (−) conditions at logarithmic phase.