Part:BBa_K108012:Experience

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Glasgow iGEM 2014 experience

We made and tested our own version of a motA biobrick. See K1463700

Our Biobrick was identical to this one (BBa_K108012) except that we used a TAA stop codon instead of the natural TGA stop codon. The natural GTG start codon is changed to ATG.

A different motA biobrick K777113 starts from an internal ATG codon in motA and therefore lacks the first 19 codons.

We characterized our motA biobrick K1463700 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

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