Difference between revisions of "Team:Austin UTexas/Parts"

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[[File:T--Austin_UTexas--Cpx_pH_Culture_Tubes_2.png|thumb|left|300px| Figure 1. Testing the CpxR Construct in pH 6-9. From left to right is Control pH 6-9 and then Experimental pH 6-9. These are showing the gradient change in expression accordingly with the change of pH due to a pH-dependent promotor compared to consistent expression accordingly with a promoter that is always "on".]]
 
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[[File:T--Austin_UTexas--pH_Dependent_Promoter.jpeg|thumb|left|500px| Figure 2. Normalized fluorescent values from CpxR construct vs control (YGCP). The fluorescence per cell count stayed generally the same throughout the range of pH while the CpxR has a clear increase in fluorescence per cell.]]
 
  
 
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<h4>CpxA-CpxR</h4>
 
<h4>CpxA-CpxR</h4>
 
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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. This system originally is a transcription factor for the virF gene, but we replaced virF with the Reporter. The original sequence was found in <i>Shigella sonnei</i>, but E. coli has a homolog of these proteins so all that is required on the construct is the appropriate prefix/suffix and CpxR binding site.
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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. This system originally is a transcription factor for the virF gene, but we replaced virF with the Reporter. The original sequence was found in <i>Shigella sonnei</i>, but E. coli has a homolog of these proteins so all that is required on the construct is the appropriate prefix/suffix and CpxR binding site. The construct can be found on the iGEM registry as: <a href="http://parts.igem.org/Part:BBa_K2097000">K2097000</a>.</p>
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Figure 1 qualitatively depicts the Control at pH 6 with more expression of the Yellow-Green Chromoprotein than the Experimental at pH 6. The pH-Dependent promoter of the Experimental group is down-regulated at pH 6 whereas the control is not. Also, there is an increase in YGCP expression between the Experiment pH 7 and pH 8 that is not seen in the Control between pH 7 and pH 8. The normalized data in Figure 2 shows the relative expression of YGCP since the qualitative data is ambiguous. The construct can be found on the iGEM registry as: <a href="http://parts.igem.org/Part:BBa_K2097000">K2097000</a>.
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Revision as of 22:43, 17 October 2016

Austin_UTexas

Registry Parts


pH Sensors

CpxA-CpxR

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. This system originally is a transcription factor for the virF gene, but we replaced virF with the Reporter. The original sequence was found in Shigella sonnei, but E. coli has a homolog of these proteins so all that is required on the construct is the appropriate prefix/suffix and CpxR binding site. The construct can be found on the iGEM registry as: K2097000.

Composite Parts

Blue Chromoprotein

K2097001 is a composite part made up of BBa_K592009 (blue chromoprotein) and BBa_K608002 (a strong RBS and promoter). The protein exhibits a strong blue/purple color when expressed. BBa_K608002 is made up of a promoter, BBa_J23104, and an RBS, BBa_B003. LINKS

Yellow-Green Chromoprotein

K2097002 is a composite part made up of BBa_K1033916 (amajLime, a yellow-green chromoprotein)and BBa_K608006 (a medium promoter and RBS). LINKS