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Team:Evry/Biology/Sensors
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| - | < | + | <h3><u><b>PCBs biosensor :</h3></u></b> |
| - | + | <br/> <div align="left"> | |
| + | <br/> <div align="justify"> | ||
| + | <br/><ul> | ||
| + | <br/><li><u><b><i>Biosensor parts</i></b></u> | ||
| + | <br/> | ||
| + | <br/>bphR2 gene, from Pseudomonas alcaligenes KF707, encodes for a prokaryotic regulatory protein. This gene is located upstream from the bph genes which are implied in the degradation of PCBs and regulates them negatively. bphR2 protein can detect PCBs when it diffuses into the cell and activate degradation genes. | ||
| + | <br/> | ||
| + | <br/>The construction is composed by a constitutive promoter <a href="http://parts.igem.org/Part:BBa_J23114">(BBa_J23114)</a>, a RBS <a href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, bphR2 gene <a href="http://parts.igem.org/Part:BBa_K1413021">(BBa_K1413021)</a>, which has been mutated because of a pstI site in its sequence, bphR1 promoter region <a href="http://parts.igem.org/Part:BBa_K1155001">(BBa_K1155001)</a>, received from Saclay’s team), RFP <a href="http://parts.igem.org/Part:BBa_E1010">(BBa_E1010)</a> and a terminator <a href="http://parts.igem.org/Part:BBa_B0015">(BBa_B0015)</a> | ||
| + | <br/><div align="center"><img src="https://static.igem.org/mediawiki/2014/a/a6/IGEM_Evry2014_Sensor_part.png" width="700px"; alt="image not found" /></div> | ||
| + | <br/> | ||
| + | <br/><li><u><b><i>How function our biosensor ?</i></b></u> | ||
| + | <br/> | ||
| + | <br/><u> In absence of PCBs : </u> | ||
| + | <br/>bphR2 <a href="http://parts.igem.org/Part:BBa_K1413021">(BBa_K1413021)</a> is bound to bphR1 promoter <a href="http://parts.igem.org/Part:BBa_K1155001">(BBa_K1155001)</a>. Transcription of RFP isn't possible. | ||
| + | <br/><div align="center"><img src="https://static.igem.org/mediawiki/2014/b/bf/IGEM_Evry2014_Absence_of_PCBs.png" width="700px"; alt="image not found" /></div> | ||
| + | <br/> | ||
| + | <br/><u> In presence of PCBs : </u> | ||
| + | <br/> When compound diffuses into the cell, it binds to bphr2 protein. This protein undergoes a conformational change and releases from the promoter that its allows the transcription of RFP. | ||
| + | <br/></ul> | ||
| + | <br/><div align="center"><img src="https://static.igem.org/mediawiki/2014/0/07/IGEM_Evry2014_Presence_of_PCBs.png" width="700px"; alt="image not found" /></div> | ||
| + | </div> | ||
| + | </div> | ||
</div> | </div> | ||
Revision as of 01:36, 11 October 2014
Biology - Classic & RNAseq-based sensors
PCBs biosensor :
- Biosensor parts
bphR2 gene, from Pseudomonas alcaligenes KF707, encodes for a prokaryotic regulatory protein. This gene is located upstream from the bph genes which are implied in the degradation of PCBs and regulates them negatively. bphR2 protein can detect PCBs when it diffuses into the cell and activate degradation genes.
The construction is composed by a constitutive promoter (BBa_J23114), a RBS (BBa_B0034), bphR2 gene (BBa_K1413021), which has been mutated because of a pstI site in its sequence, bphR1 promoter region (BBa_K1155001), received from Saclay’s team), RFP (BBa_E1010) and a terminator (BBa_B0015)
- How function our biosensor ?
In absence of PCBs :
bphR2 (BBa_K1413021) is bound to bphR1 promoter (BBa_K1155001). Transcription of RFP isn't possible.
In presence of PCBs :
When compound diffuses into the cell, it binds to bphr2 protein. This protein undergoes a conformational change and releases from the promoter that its allows the transcription of RFP.
