Reporter Project

From 2011.igem.org

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=== The Reporter ===
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<h2>The Reporter</h2>
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<p> Our reporter system focused on modifying the part created by Imperial College London iGEM 2010 for their fast reporter system. Imperial’s quick acting response module used the transcription of TEV protease to cleave the bond between the GFP, C23O fusion protein. This unbound form of C230 could then form a tetramer and convert the colorless substrate catechol into a yellow product. The details on their part can be found <a href="http://partsregistry.org/Part:BBa_K316007">here</a>.  
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|Our reporter system will use a modified version of Imperial College London iGEM 2010 fast reporter system. Imperial’s quick acting response module used the transcription of TEV protease to cleave the bond between the GFP, C23O fusion protein. This unbound form of C230 could then form a tetramer and convert the colorless substrate catechol into a yellow product.
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<h2>Our modifications and expansions of part K316007: Recombinant GFP-XylE protein</h2>
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<p>We have cloned and created a catalog of the individual fusion parts included in the fast acting reporter, each with standard 25 prefix and suffix. These parts can be quickly assembled using the Freiburg BioBricking Assembly technique,   and recombined to produce and experiment with new variations of the recombinant fusion protein. New variations of this part can be created, optimized, and applied to many different situations requiring the use of a rapid reporter in a "in vivo" system. For example in our proposed bacterial dosimeter device.
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|Using the Freiburg BioBricking Assembly technique, we plan to produce a miniature catalog of fast acting reporters while reducing the leakiness of Imperial’s construct. To achieve the latter, we plan to link a second GFP to the C-terminus of C230; this new linkage would be cleavable by RecA.  
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Along these lines our team has designed our own iterations and improvements to the fast reporter protein. In particular, we focused on reducing any leakiness of Imperial’s construct (see our experimental data section for information on our assessment of the original part. To achieve this we plan to link a second GFP to the C-terminus of C230; this new linkage would be cleavable by RecA and create a fast reporter system specifically customized for our biosensor system.</p>
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<p> Other proposed modifications by our team included the use of new fusion protein linkers and new enzyme subunits, specifically the B-glucuronidase and B-galactose reporters. These two possible enzymes could be made to produce rapidly a blue color product from substrates that are slightly less toxic to cells then catechol.</p>
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Revision as of 18:08, 27 September 2011

RecA Project Sensor Project Reporter Project

The Reporter

Our reporter system focused on modifying the part created by Imperial College London iGEM 2010 for their fast reporter system. Imperial’s quick acting response module used the transcription of TEV protease to cleave the bond between the GFP, C23O fusion protein. This unbound form of C230 could then form a tetramer and convert the colorless substrate catechol into a yellow product. The details on their part can be found here.

Our modifications and expansions of part K316007: Recombinant GFP-XylE protein

We have cloned and created a catalog of the individual fusion parts included in the fast acting reporter, each with standard 25 prefix and suffix. These parts can be quickly assembled using the Freiburg BioBricking Assembly technique, and recombined to produce and experiment with new variations of the recombinant fusion protein. New variations of this part can be created, optimized, and applied to many different situations requiring the use of a rapid reporter in a "in vivo" system. For example in our proposed bacterial dosimeter device.

Along these lines our team has designed our own iterations and improvements to the fast reporter protein. In particular, we focused on reducing any leakiness of Imperial’s construct (see our experimental data section for information on our assessment of the original part. To achieve this we plan to link a second GFP to the C-terminus of C230; this new linkage would be cleavable by RecA and create a fast reporter system specifically customized for our biosensor system.




Other proposed modifications by our team included the use of new fusion protein linkers and new enzyme subunits, specifically the B-glucuronidase and B-galactose reporters. These two possible enzymes could be made to produce rapidly a blue color product from substrates that are slightly less toxic to cells then catechol.


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