Team:Aachen/Project/FRET Reporter
From 2014.igem.org
(→A Fluorescence Answer Faster Than Expression) |
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==Producing a GFP_Reach Fusion Protein== | ==Producing a GFP_Reach Fusion Protein== | ||
- | In our project, we reproduced the REACh1 and REACh2 proteins by subjecting an RFC-25 compatible version of the BioBrick [http://parts.igem.org/Part:BBa_E0030 E0030] (EYFP) to a '''QuikChange mutation'''. Subsequently, we fused each REACh protein with ''' | + | In our project, we reproduced the REACh1 and REACh2 proteins by subjecting an RFC-25 compatible version of the BioBrick [http://parts.igem.org/Part:BBa_E0030 E0030] (EYFP) to a '''QuikChange mutation'''. Subsequently, we fused each REACh protein with '''GFP (mut3b)''' which is available as BioBrick [http://parts.igem.org/Part:BBa_E0040 E0040]. The protein complex was linked via a '''protease cleavage site'''. Therefore, when GFP is connected to either REACh quencher, GFP will absorb light but the energy will be transferred to REACh via FRET and then emitted in the form of heat. However, when the complex is cleaved by a protease, REACh will be separated from GFP. The latter will then be able to absorb and emit light as usual. |
''Add: [Fig. 4: DNA scheme of BioBricks, and scheme of fusion protein]'' | ''Add: [Fig. 4: DNA scheme of BioBricks, and scheme of fusion protein]'' |
Revision as of 13:02, 7 October 2014
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