Speeding Up Device
|Speeding Up Device makes E.coli to move in higher speed than usual. E.coli have HNS protein gene in their genome. HNS is an Escherichia coli nucleoid protein known only to function as a modulator of gene expression. Gina M. Donato said that specific single amino acid substitutions in H-NS caused an approximately 50% increase in flagellum rotational speed (*1). So we have E.coli to express HNS mutant T108I gene with plasmid.|
|We designed the part BBa_K543003 HNS mutant T108I protein generator. With this part, chassis would produce HNS mutant T108I protein and get hight speed move.|
Swarm plate assay*1|
Fresh colonies from strains carrying the indicated alleles were inoculated onto semi-solid agar plates and grown at 30 °C. Growth is measured as the diameter of the bacterial swarm over several time points. Bacteria with swarm diameters under 10 mm at the end of 17 h incubation were considered non-motile.
fig.1 swarm assay
Succeeded in clonning of HNS wild type protein and registried it for BBa_K543000.
402 guanine is mutated into cytosine in this part to prevent XbaⅠ digesting it.
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Gina M. Donato and Thomas H. Kawula 1998, Enhanced Binding of Altered H-NS Protein to Flagellar Rotor Protein FliG Causes Increased Flagellar Rotational Speed and Hypermotility in Escherichia coli