Here, we had a look if the protein produced by our biobrick was well traducted et abled to recover swing in a FliC deficient strain. Cells were ensemenced using a tooth pic and incubate at 37☌. We tested 3 background: W3110 (down left) knockout W3110 fliC mutant(down right) and knockout W3110 fliC mutant complemented with BBa_K1951008(up). Escherichia coli W3110 strain has been used as a wild type because of its good swimming capacity. We investigated here if swimming was recovered by a knockout FliC strain complementation with our biobrick on soft LB agar gelose. The strong J23100 promoter in this result contained a mutation rendering it inactive see K1463604.įor more information on the biobrick and methods used go to The error bars indicate the range or results obtained in two repeats of the experiment. The diameter of swimming in a 16 hour swarm assay at 37 degrees is shown. Plasmids containing these constructs were used to complement a chromosomal fliC mutation. (G) DS941 ΔfliC + J23106-fliC(1), (H) DS941 ΔfliC + J23106-fliC(2)įigure 2 - FliC Motility Histogram The promoters indicated in the histogram were used to drive the fliC biobrick K1463600 with the RBS B0034. (C) DS941 ΔfliC + pSB1C3 fliC (no promoter), (D) DS941 ΔfliC + J23100 (mutant promoter) fliC, When used in conjunction with various promoters (J23106 and J23116) and B0032 ribosome binding site, this biobrick was shown to restore swimming in fliC knockout strains (See Figures 1 & 2).įlic Motility Swarm Assay Test from Glasgow team 2014 A promoter is necessary to drive fliC expression. In a fliC knockout strain, this biobrick alone was not enough to restore swimming. See Fig 1C for motility swarm assay results showing the function of this biobrick. coli and is thus fundamental to motility. FliC encodes the major flagellar protein of E.
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