Fig. 8.
Validation of a TSS and a promoter of the newly annotated and proteomics-supported gene bll0506_ISGA. a The bll0506_ISGA locus with cDNA reads and mapped TSSs (red flexed arrows) and promoters (red squares). The analyzed TSS Bja_TSS_6328 with a low peak height and a low-score (no. 6 in Fig. 5), and its RpoD-like promoter are in dark red; non-analyzed TSSs and promoter are in light red. The bll0506_ISGA mRNA is shown as a bar with differently colored regions: grey, the annotated bll0506 ORF; dark red, the here verified 5′-UTR; light red, a proposed upstream part of a longer 5′-UTR. b The wild type sequence upstream of Bja_TSS_6328 containing the RpoD-like promoter (wtP6328) and the mutated version mutP6328. Bases subjected to mutagenesis are underlined. For other descriptions see the legend of Fig. 4d. c Beta-galactosidase activity measurements in B. japonicum. The wild-type and mutated versions of the 41 nt-region upstream of Bja_TSS_6328 (see panel b) were transcriptionally fused to the lac operon. Shown are results from three independent experiments with technical duplicates and error bars depicting the standard deviation