Team:Aachen/Parts
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AZimmermann (Talk | contribs) (→RFC [25] - compatible dark quencher based on K1319000 (E0030)) |
AZimmermann (Talk | contribs) (→RFC [25] - compatible dark quencher based on K1319000 (E0030)) |
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<span class="anchor" id="partsK1319001"></span> | <span class="anchor" id="partsK1319001"></span> | ||
- | ===RFC [25] - compatible dark quencher based on K1319000 (E0030)=== | + | ===RFC [25] - compatible dark quencher based on K1319000 (E0030) called REACh 1=== |
This part is an RFC [25] dark quencher that is based upon K1319000 (the RFC [25] version of E0030/EYFP). | This part is an RFC [25] dark quencher that is based upon K1319000 (the RFC [25] version of E0030/EYFP). | ||
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*Y145W | *Y145W | ||
- | This protein is designed to be a dark quencher for GFP ([http://parts.igem.org/Part:BBa_E0040 E0040]) in a FRET system. When used in a fusion protein with GFP it reduces the observed fluorescence of GFP drastically. In the biobrick [http://parts.igem.org/Part:BBa_K1319013 K1319013] this is realised and the proteins are fused with the linker [http://parts.igem.org/Part:BBa_K1319016 K1319016] which includes a specific TEV protease cleavage site. | + | This protein is designed to be a dark quencher for GFP ([http://parts.igem.org/Part:BBa_E0040 E0040]) in a FRET system. When used in a fusion protein with GFP it reduces the observed fluorescence of GFP drastically. In the biobrick [http://parts.igem.org/Part:BBa_K1319013 K1319013] this is realised and the proteins are fused with the linker [http://parts.igem.org/Part:BBa_K1319016 K1319016] which includes a specific TEV protease (available as [http://parts.igem.org/Part:BBa_K1319004 K1319004]) cleavage site. The fusion of the proteins bring GFP and REACh 1 in proximity to each other which allows GFP and REACh 1 to act as donors and acceptors in a FRET (Förster Energy Transfer System) system. GFPs emission energy is thereby taken up by REACh 1 and released as thermal energy instead of visible light. This eliminates the GFP fluorescence and allows for a release of a strong fluorescence signal if a TEV protease is expressed and the linker is cut. This separates GFP and REACh 1 cancelling the FRET interaction and providing a GFP fluorescence response. |
{{Team:Aachen/BlockSeparator}} | {{Team:Aachen/BlockSeparator}} |
Revision as of 20:19, 16 October 2014
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