Composite

Part:BBa_K1615097

Designed by: Brooke Rothschild-Mancinelli   Group: iGEM15_Edinburgh   (2015-08-31)

CBDclos fused to mRFP in RFC25

mRFP (BBa_K1615089) fused to the C terminal of CBDclos (BBa_K1321002). If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.

CBD RFP folding.jpg

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.

BBa K1615097.jpg

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. The results show a very low binding affinity of CBDclos.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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