FIG. 6.

Analysis of the N. gonorrhoeae lst promoter. (A) Primer extension analysis. Total RNA (20 μg) was isolated from N. gonorrhoeae strain F62, and primer extension analysis was performed using primer PE1. The extension product along with a sequencing reaction with the same primer was resolved by 6% PAGE with 8 M urea. The arrow points to the transcriptional start site. (B) RNase protection assay. ³²P-labeled RNA probe was hybridized with total RNA from N. gonorrhoeae strains F62 and FA1090 or yeast RNA (negative control) and digested with a 1:100 dilution of RNase A/T₁ mix. The protected fragments were resolved along with a DNA size marker by 6% PAGE with 8 M urea. (C) The 5′lst upstream sequence of N. gonorrhoeae F62. The −10 and −35 sequences of the promoter are boldface and underlined, and their positions relative to the translational start site are given. The transcriptional start site (tsp) is marked by an arrow; the putative Shine-Dalgarno (SD) sequence and translational start site are in boldface.