Difference between revisions of "Team:Edinburgh/Composite Part"
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| − | <a data-toggle="collapse" data-parent="#accordion" href="# | + | <a data-toggle="collapse" data-parent="#accordion" href="#collapseFour">mRFP fused to CBDcenA+Linker in RFC25 (BBa_K16150092)</a> |
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| − | <a data-toggle="collapse" data-parent="#accordion" href="# | + | <a data-toggle="collapse" data-parent="#accordion" href="#collapseFive">Linker +dCBD fused to mRFP in RFC25 (BBa_K1615093) |
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Revision as of 19:08, 20 November 2015
mRFP
mRFP (BBa_K1615089) driven by the IPTG inducible PLlac 0-1 promoter (BBa_K314103). This allows the mRFP gene to express mRFP when induced with IPTG.
mRFP (BBa_K1615089) fused to the C terminal of CBDcenA + C-terminal linker (BBa_K1321339) in RFC25. If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.
To test the RFP folding when fused to a CBD we placed 200µL of crude cell lysate in a 96 well plate and observed the fluorescence under a blue light transilluminator.
mRFP (BBa_K1615089) fused to the N terminal of CBDcenA + C terminal linker (BBa_K1321339) in RFC25. If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.
mRFP (BBa_K1615089) fused to the N terminal of CBDcenA + C terminal linker (BBa_K1321339) in RFC25. If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.
We tested the dissociation of this part on Whatman 54 using isostandard chads. This involved incubating the chads in the crude cell lysate for 10 minutes and then washing them with PBS for various periods time to test the binding affinity of this CBD.
We tested the dissociation of this part on Whatman 54 using isostandard chads. This involved incubating the chads in the crude cell lysate for 10 minutes and then washing them with PBS for various periods time to test the binding affinity of this CBD.
mRFP (BBa_K1615089) fused to the C terminal of dCBD + N terminal linker (BBa_K1321340). If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.
To test the RFP folding when fused to a CBD we placed 200µL of crude cell lysate in a 96 well plate and observed the fluorescence under a blue light transilluminator.