Difference between revisions of "Part:BBa K2097000"
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| + | <partinfo>BBa_K2097000 SequenceAndFeatures</partinfo> | ||
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| + | ===Design Notes=== | ||
| + | The sequence directly from the papers - "Involvement of cpxA, a Sensor of a Two-Component Regulatory System, in the pH-Dependent Regulation of Expression of Shigella sonnei virF Gene" (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC177285/pdf/1775062.pdf) and "Identification of cpxR as a Positive Regulator Essential for Expression of the Shigella sonnei virF Gene" (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC107317/pdf/jb003522.pdf) - was not used. We used the loci detailed in the papers and found the CpxR binding sequence in the GenBank to construct the sequenced used in the experiments. | ||
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Revision as of 17:57, 20 October 2016
CpxR binding site attached to a yellow-green color protein (YGCP) acts as a neutral pH indicator.
CpxA-CpxR is a two-component mechanism that is activated at pH 7.4 and repressed at pH 6.0. CpxA is an intermembrane protein that autophosphorylates at a certain external pH, CpxR (a kinase) then gets phosphorylated by CpxA and acts as a transcription factor for the downstream gene, YGCP in this case. This system originally is a transcription factor for the virF gene, but virF was replaced with a reporter. The original sequence was found in Shigella sonnei, but E. coli has a homolog of these proteins so only the appropriate BioBrick prefix/suffix and CpxR binding site were required. Used BBa_K2097002 as the control series.
Usage and Biology
Figure 1.