Part:BBa_K777113:Experience

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BBa_K777113 Glasgow team 2014

Glasgow iGEM 2014 experience We made our own version of a motA biobrick. See K1463700 Changes we made : 1. The natural motA gene starts at a GTG codon. This motA biobrick (K777113) started at the first ATG codon within motA, not the natural GTG, and therefore started at the wrong start codon 58 bp into the natural full length motA. This motA biobrick (K777113) therefore encodes a protein missing the first 19 amino acids. Our motA has the correct start as annotated on the E. coli genome sequence. 2. We changed the wild-type GTG start codon to ATG to meet registry requirements. 3. We changed the stop codon was changed to the preferred TAA. We characterized our motA biobrick using soft agar swimming assays. We found that a composite biobrick containing motA and motB restored swimming to our motA knock-out mutant. The extent of complementation depended on the strength of promoter used. Fig. 1: DS941 ΔmotA E. coli carrying plasmids with the indicated biobricks were tested for mobility on swarm plates. Growth diameter of swarm assay grown for 16 hours at 37 degrees C. The first column shows DS941, a non knock-out strain as control. The other columns show the motA knockout mutant (DS941 ΔmotA) with plasmids carrying motA and motB driven by different strength constitutive promoters. For more details see K1463700 UNIQ72d729ed8a870c56-partinfo-00000002-QINU