Skip to main content
. 2000 Aug 15;97(18):10220–10224. doi: 10.1073/pnas.170219997

Figure 1.

Figure 1

Ectopic expression of OmpU and OmpT. (A) The ompT promoter fragment corresponding to −429 to +104 with respect to the transcription start site was fused to the ompU ORF to form Tp-U (pKEK256), which allows ectopic expression of OmpU in a toxR− strain. The ompU promoter fragment corresponding to −674 to +149 with respect to the transcription start site was fused to the ompT ORF to form Up-T (pKEK257), which allows ectopic expression of OmpT in a toxR+ strain. Translational fusions were facilitated by the incorporation of an NdeI site (CATATG) at the initiating Met codon of each ORF. (B) V. cholerae strains O395 (toxR+; lane 1); KKV61 (ΔtoxR; lane 2); KKV780 (ΔompU) carrying either plasmid pWSK30 (vector, lane 3); pKEK253 (Up-U, lane 4); or pKEK257 (Up-T, lane 5); and KKV 804 (ΔompT ΔtoxR) carrying either plasmid pWSK30 (vector, lane 6), pKEK255 (Tp-T, lane 7), or pKEK256 (Tp-U, lane 8). Whole-cell lysates were matched by OD600 and separated on 10% SDS/PAGE, then stained with Coomassie blue; left-hand lane contains molecular mass standards (in kDa), and OmpT and OmpU bands are designated by arrows. (C) The samples of lanes 1–8 were subjected to Western analysis by using rabbit polyclonal antisera against OmpT (α-OmpT, Upper) and OmpU (α-OmpU, Lower).