Difference between revisions of "Part:BBa K1497019"
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FdeR is a homo dimeric protein from ''Herbaspirillum seropedicae''. In present of naringenin, FdeR activates the specific promotor region upstream of the fdeR region and allow a strong gene expression. | FdeR is a homo dimeric protein from ''Herbaspirillum seropedicae''. In present of naringenin, FdeR activates the specific promotor region upstream of the fdeR region and allow a strong gene expression. | ||
− | + | [[File:Homology_model_of_FdeR.png|300px|thumb|right|Homology model of FdeR. PDB: 2ESN was used as template. ]] | |
Based on this reporter protein and 3 different fluorescence proteins, we designed 3 new biosensors for in vivo detection and determination of naringenin. | Based on this reporter protein and 3 different fluorescence proteins, we designed 3 new biosensors for in vivo detection and determination of naringenin. | ||
− | |||
− | ===Usage and Biology== | + | |
+ | ===Usage and Biology=== | ||
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<html> | <html> | ||
<BLOCKQUOTE> | <BLOCKQUOTE> | ||
− | <li>with GFP response | + | <li>with GFP response use <a href="/Part:BBa_K1497020">BBa_K1497020</a></li> |
− | <li>with mKate response | + | <li>with mKate response use <a href="/Part:BBa_K1497021">BBa_K1497021</a></li> |
− | <li>with CFP response | + | <li>with CFP response use <a href="/Part:BBa_K1497022">BBa_K1497022</a></li> |
</BLOCKQUOTE> | </BLOCKQUOTE> | ||
</html> | </html> | ||
+ | <html> | ||
+ | <div align="center"> | ||
+ | <table class="MsoTableGrid" | ||
+ | style="border: medium none ; border-collapse: collapse; text-align: left; margin-left: auto; margin-right: auto;" | ||
+ | border="0" cellpadding="0" cellspacing="0"> | ||
+ | <tbody> | ||
+ | <tr style="height: 214.9pt;"> | ||
+ | <td | ||
+ | style="padding: 0cm 5.4pt; vertical-align: top; width: 236.7pt; height: 214.9pt;"> | ||
+ | <img | ||
+ | style="width: 638px; height: 308px;" alt="" | ||
+ | src="https://static.igem.org/mediawiki/2014/3/38/Naringeninsensensorschemagr%C3%BCn.png"></p> | ||
+ | <br> | ||
+ | <p class="MsoCaption" align="text-align:justify"><span lang="EN-US"><b>Figure 2</b></span></a><span lang="EN-US"> | ||
+ | Flow chart of the FdeR activated gfp expression. The constitutive expression of fdeR forms the homodimeric FdeR protein. In present of naringenin, , naringenin molecules bind to the FdeR protein and operate a conformational change of the homodimeric FdeR structure. This conformational change activates FdeR, which is now enabled to bind to the uncharacterized promotor domain. Binding to the promotor domain induces expression of genes, which are cloned behind fdeR promtor region. </span></p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tbody> | ||
+ | </table> | ||
+ | </div> | ||
+ | </html> | ||
− | You can create your own naringenin sensor or your own naringenin dependent gene expression device as well. For these reasons use | + | You can create your own naringenin sensor or your own naringenin dependent gene expression device as well. For these reasons use the Biobrick <html><a href="/Part:BBa_K1497019">K1497019</a></html> and clone your parts of interest (without RBS!) behind the device. |
Revision as of 22:38, 9 October 2014
fdeR (Naringenin binding protein) and fde operon regulatory domain
FdeR is a homo dimeric protein from Herbaspirillum seropedicae. In present of naringenin, FdeR activates the specific promotor region upstream of the fdeR region and allow a strong gene expression.
Based on this reporter protein and 3 different fluorescence proteins, we designed 3 new biosensors for in vivo detection and determination of naringenin.
Usage and Biology
You can use the reporters for measuring naringenin concentrations in your samples. Depending on which fluorophor you want to detect, you can use one of three biosensors:
with GFP response use BBa_K1497020 with mKate response use BBa_K1497021 with CFP response use BBa_K1497022
Figure 2 Flow chart of the FdeR activated gfp expression. The constitutive expression of fdeR forms the homodimeric FdeR protein. In present of naringenin, , naringenin molecules bind to the FdeR protein and operate a conformational change of the homodimeric FdeR structure. This conformational change activates FdeR, which is now enabled to bind to the uncharacterized promotor domain. Binding to the promotor domain induces expression of genes, which are cloned behind fdeR promtor region. |
You can create your own naringenin sensor or your own naringenin dependent gene expression device as well. For these reasons use the Biobrick K1497019 and clone your parts of interest (without RBS!) behind the device.
Functional Parameters
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 95
Illegal NgoMIV site found at 452
Illegal NgoMIV site found at 543
Illegal NgoMIV site found at 555 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 271