FIG 2.

RT-PCR analysis of ech expression. The expression of ech at different stages of the fermentation was analyzed by RT-PCR. Samples were taken prior to feeding with ferulic acid and after 7 h of biotransformation. RNA isolation, preparation of cDNA, and the final PCR were performed as described in Materials and Methods. The resulting fragments were analyzed in a 1% agarose gel. Lanes M, molecular size markers (GeneRuler 1-kb DNA ladder; Fermentas GmbH, St. Leon-Rot, Germany); lanes 1 to 5, amplification products obtained using the oligonucleotides ech_RT_for and ech_RT_rev and different templates; lane 1, cDNA prepared from a sample prior to feeding; lane 2, RNA control for the sample in lane 1; lane 3, cDNA prepared from a sample during feeding; lane 4, RNA control for the sample in lane 3; lane 5, DNA control for PCR. Arrow, expected internal 529-bp fragment of ech.