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. 2008 Jan 18;190(6):2128–2137. doi: 10.1128/JB.01746-07

FIG. 5.

FIG. 5.

LPS profiles and protein expression for Wzz mutants with single amino acids replacements. LPS was prepared from cultures with 0.2% (wt/vol) arabinose. Samples were analyzed as described in the legend to Fig. 3. All the plasmids were introduced into EVV33(pMF19) by transformation. (A) Lane 1, pBADFlag; lane 2, pEH1 (Wzz); lane 3, pEH6 (Wzz-I); lane 4, pEH1-L118F; lane 5, pEH1-L122F; lane 6, pEH1-L122A; lane 7, pEH1-F115A; lane 8, pEH1-F115F; lane 9, pEH1-A116F. (B) Lane 1, pBADFlag; lane 2, pEH1 (Wzz); lane 3, pEH4 (Wzz-II); lane 4, pEH1-A156F; lane 5, pEH1-A156L; lane 6, pEH1-L160F. (C) Twenty micrograms of total membranes was separated on a 14% SDS-PAGE gel and transferred to a nitrocellulose membrane, and the blot was incubated with anti-Flag monoclonal antibodies. The specific bands were detected by fluorescence using Alexa Fluor 680 anti-mouse IgG antibodies. Lane M, broad-range prestained SDS-PAGE standard (Bio-Rad); lane 1, pBADFlag; lane 2, pEH1 (Wzz); lane 3, pEH6 (Wzz-I); lane 4, pEH1-L118F; lane 5, pEH1-L122F; lane 6, pEH1-L122A; lane 7, pEH1-F115A; lane 8, pEH1-F115F; lane 9, pEH1-A116F; lane 10, pEH4 (Wzz-II); lane 11, pEH1-A156F; lane 12, pEH1-A156L; lane 13, pEH1-L160F.