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. 2011 Aug;193(15):3710–3721. doi: 10.1128/JB.00059-11

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Fig. 5. — Effect of reciprocal single- and multiple-domain swapping of small regions of WzzB from S. flexneri and S. Typhimurium. (A) silver-stained polyacrylamide gel showing the O-antigen LPS profile of E. coli EVV33 (pMF19) expressing different forms of WzzB (Table 1). The designations T7 to T13 indicate that the chimera contains one segment from WzzB_ST in the WzzB_SF background, while T14 to T17 denote two or more segments of WzzB_ST in the WzzB_SF background. The designations F7 to F13 indicate that the chimera contains one segment from WzzB_SF in the WzzB_ST background. F14 to F17 denotes two or more segments of WzzB_SF in the WzzB_ST background. Lanes 1 and 11, parental WzzB_SF; lane 2, WzzB_SFV67-L95>A67-F95WzzB_ST (WzzB-SF-T7); lane 3, WzzB_SFQ202-I274>E202-V274WzzB_ST (WzzB-SF-T8); lanes 4 and 17, WzzB_SFQ202-A253>E202-N253WzzB_ST (WzzB-SF-T9); lane 5, WzzB_SFQ202-K214>E202-Q214WzzB_ST (WzzB-SF-T10); lane 6, WzzB_SFN260-I274>T260-V274WzzB_ST (WzzB-SF-T12); lane 7, WzzB_SFF269-I274>V269-V274WzzB_ST (WzzB-SF-T13); lane 8, WzzB_STA67-F95>V67-L95WzzB_SF (WzzB-ST-F7); lane 9, pBAD vector control; lane 10, parental WzzB_ST; lane 12, WzzB_SFV217-E237>I217-K237WzzB_ST (WzzB-SF-T11); lane 13, WzzB_SFQ202-K214>E202-Q214/F269-I274>V269-V274WzzB_ST (WzzB-SF-T14); lane 14, WzzB_SFV67-L95>A67:F95/F269-I274>V269-V274 WzzB_ST (WzzB-SF-T15); lane 15, WzzB_SFV67-L95>A67-F95/Q202-K214>E202-Q214WzzB_ST (WzzB-SF-T16); and lane 16, WzzB_SFV67-L95>A67:F95/Q202-K214>E202-Q214/F269-I274>V269-V274WzzB_ST (WzzB-SF-T17). (B) Silver-stained polyacrylamide gel showing the O-antigen LPS profile of E. coli EVV33 (pMF19) expressing various recombinant WzzB forms (Table 1). Lane 1, WzzB_STE202-V274>Q202-I274WzzB_SF (WzzB-ST-F8); lane 2, WzzB_STE202-N253>Q202-A253 WzzB_SF (WzzB-ST-F9); lane 3, WzzB_STE202-Q214>Q202-K214WzzB_SF (WzzB-ST-F10); lane 4, WzzB_STI217-K237>I217-E237WzzB_SF (WzzB-ST-F11); lane 5, WzzB_STT260-V274>N260:I274WzzB_SF (WzzB-ST-F12); lane 6, WzzB_STV269-V274>F269-I274WzzB_SF (WzzB-ST-F13); lanes 7 and 9, parental WzzB_ST; lane 8, pBAD vector control; lane 10, parental WzzB_SF; 11, WzzB_STE202-Q214>Q202-K214/T260-V274>N260-I274WzzB_SF (WzzB-ST-F14); lane 12, WzzB_STA67-F95>V67-I95/E202-Q214>Q202-K214WzzB_SF (WzzB-ST-F15); lane 13, WzzB_STE202-Q214>Q202-K214/V269-V274>F269-I274WzzB_SF (WzzB-ST-F16); and lane 14, WzzB_STA67-F95>V67-I95/V269-V274>F269-I274WzzB_SF (WzzB-ST-F17. Dotted lines indicate the average modality for the E. coli K-12 (O16) LPS OAg molecules, as determined by WzzB_ST (T), WzzB_ST-like (II), WzzB_SF-like (I), and WzzB_SF (F) proteins. (C and D) Graphical representation of the changes in modality observed for hybrids in panel A, lanes 2 to 7 and 12 as well as lanes 13 to 16, respectively. (E and F) Graphical representation of the changes in modality observed for hybrids in panels A (lane 8) and B (lanes 1 to 6) as well as in panel B, lanes 11 to 14, respectively. Hybrid polypeptides are plotted according to the residue number (on the x axis) and the average modality (OAg repeat number; on the y axis). Thick red segments indicate the regions derived from WzzB_ST, while thick blue segments indicate WzzB_SF regions. A linear representation of the structural map of WzzB and the location of the hybrid endpoints are also indicated in panels C through F.