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. 2007 Jan 19;189(7):2618–2628. doi: 10.1128/JB.01905-06

FIG. 6.

FIG. 6.

(A) Expression of WecAFLAG-7×His. Total membranes were prepared from strain MV501 (VW187 wecA::Tn10) transformed with various plasmids containing several versions of wecA (Table 1). Lane 1, WecAFLAG-7×His, pJL1; lane 2, WecAFLAG-7×His-Cys, cysteineless version of WecA encoded by pJL7; lane 3, WecAFLAG-5×His, pKV1; lane 4, vector control, pBAD-His. Each lane contained 4 μg of protein. Membranes were transferred to a nitrocellulose membrane and reacted with anti-FLAG monoclonal antibodies. The molecular mass standards were myosin (206 kDa), β-galactosidase (119 kDa), bovine serum albumin (91 kDa), ovalbumin (51 kDa), and carbonic anhydrase (34.7 kDa). (B) Complementation of O7 LPS synthesis in strain MV501 (VW187 wecA::Tn10). LPS samples were obtained from E. coli MV501 (VW187 wecA::Tn10) transformed with various plasmids. Lane 1, vector control, pBAD-His; lane 2, WecAFLAG-5×His, pKV1; lane 3, WecAFLAG-7×His-Cys, cysteineless version of WecA encoded by pJL7. LPS was separated by Tricine-SDS-PAGE, which was followed by silver staining. Loading was normalized by determining the amount of KDO in the inner core.