FIG. 4.

Identification of the transcriptional start site for OpdA. Strain TN4465 containing pCM138 was grown overnight at 30°C, diluted 1:100, and incubated at 30°C to an optical density at 600 nm of 0.4. The culture was then shifted to 42°C for 10 min, and total RNA was isolated by using Qiagen's RNeasy Mini kit. A 25-nt primer complementary to nt 603 to 627 of GenBank sequence accession no. M84574 was used for extension, and the product is shown in lane PE. Lanes A, C, G, and T contain sequence reactions generated with the same primer. Radioactivity was detected by autoradiography and, in addition, by the use of a Molecular Dynamics Storm 860 PhosphorImager. An asterisk indicates the transcription start in the DNA sequence shown on the left.