Difference between revisions of "Team:WLC-Milwaukee/Parts"

Of all the tolC proteins we used in our project, three were ultimately submitted to the BioBrick registry. These submitted parts were tolC homologs from Yersenia pestis Angola (BBa_K1683000), Proteus mirabilis (BBa_K1683002), and Pseudomonas aeruginosa PA01 (oprM) (BBa_K168004). These proteins were submitted as a part of larger constructs which we used to express usable tolC molecules in our chassis organism, ΔtolC E. coli. These constructs consist of 4 major components. First is the pBAD promoter, an arabinose-induced strong E. coli promoter. Second is the strong E. coli ribosomal binding site (originally from BBa_B0034). Third is the E. coli tolC signaling sequence; this forms a chimeric protein with the tolC or tolC homolog of interest. See Project > Solution > TolC for more information on TolC.

These parts were characterized by western blotting and through Kirby-Bauer antibiotic assays. More detailed analysis is found on the Documentation page’s Results section. The important snippets relating to the submitted parts have been extracted for this page.