Fig. 3. The espADB mRNAs have heterogeneous 5’ ends.

Total RNA was extracted from cells collected from a DMEM culture of the Sakai strain. RPA was performed with probe 1, but in this case the probe was unlabeled. The RPA reaction products were resolved in a denaturing polyacrylamide gel, and transferred to a membrane. To detect the products, the membrane was cut into two halves. One half was hybridized with a DNA oligomer specific for sepL and the other was hybridized with a DNA oligomer specific for espA. Both oligomers were labeled with [γ-³²P]ATP at their 5’ ends (represented as lines with a star). On the right side of the figure, a scheme represents the probe and protected fragments detected in this assay (IR, intergenic region).