Team:WLC-Milwaukee/Description

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Expression of Salmonella and Vibrio cholerae TolC protein – A western blot was performed using α-TolC specific against Escherichia coli antibodies. These antibodies also recognize the TolC proteins from Salmonella and Vibrio cholerae. Lane 1 contains the protein ladder (NEB Prestained protein ladder, broad range, NEB #7702) which demonstrates the sizes of the TolC proteins displayed in the western blot are correct according to the predicted molecular weights for each. The next two lanes are wild-type E. coli (BW25113) with the chromosomal tolC gene (Lane 2) or without (Lane 3). The following three lanes are the E. coli strains containing the tolC deletion and the pUC57 plasmid with the arabinose promoter alone (Lane 4), the arabinose promoter followed by the Salmonella tolC gene (Lane 5), or the arabinose promoter followed by the Vibrio cholerae tolC gene (Lane 6). No arabinose was added to the cultures used for the samples in Lanes 4, 5, and 6. Lanes 7, 8, and 9 were the same strains as the previous three lanes except that 0.1% arabinose was added to each culture.