Part:BBa_K1321346:Experience

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Applications of BBa_K1321346

As part of our project we carried out an assay to determine the relative binding ability of CBD-sfGFP fusions to bacterial cellulose.

Our first assay was performed to determine the relative strengths of various CBDs’ binding to bacterial cellulose. These were represented by the percentage fluorescence left from CBDs fused to (sfGFP (RFC25)) bound to bacterial cellulose discs, when subjected to various washes. Full protocol can be found ([http://2014.igem.org/Team:Imperial/Protocols here])

In the same assay, results suggested that dCBD, CBDcex, CBDcipA, and C-terminal CBDclos (all fused to sfGFP) had greater ability to bind bacterial cellulose than N-terminal CBDclos. Three washes with dH2O, PBS and 5% BSA were carried out to monitor binding strength. It was determined that the fusion with CBDclos was, on average, our fourth strongest CBD, in comparison other CBDs tested.

Figure 1 - CBD-sfGFP binding strength after three dH2O washes

Figure 2 - CBD-sfGFP binding strength after three 70%Ethanol washes

Figure 3 - CBD-sfGFP binding strength after three PBS washes

Figure 4 - CBD-sfGFP binding strength after three BSA washes

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