Difference between revisions of "Part:BBa K1321343:Experience"

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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
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===Applications of BBa_K1321343===
 
===Applications of BBa_K1321343===
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As part of our project we carried out an assay to determine the relative binding ability of CBD-sfGFP and sfGFP fusions to bacterial cellulose. 
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For reference, the cellulose binding domain binding capability of CBDclos (C-terminally fused) to bacterial cellulose was measured relative to other cellulose binding domains when fused to sfGFP, the data for which can be seen [https://parts.igem.org/Part:BBa_K1321341 here] - K1321341.
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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 ([https://parts.igem.org/Part:BBa_K1321337 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]) 
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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 N-terminal CBDclos was, on average, our fourth strongest CBD, in comparison other CBDs tested.
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[[File:IC14_-_dH2Obplot1.png|600px|thumb|left|Figure 1 - CBD binding strength after three dH2O washes. ]]
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[[File:IC14-EtOHbplot1.png|600px|thumb|left|Figure 2 - CBD binding strength after three 70%Ethanol washes]]
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[[File:IC14-PBSbplot1.png|600px|thumb|left|Figure 3 - CBD binding strength after three PBS washes]]
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[[File:IC14-BSAbplot1.png|600px|thumb|left|Figure 4 - CBD binding strength after three BSA washes]]
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===User Reviews===
 
===User Reviews===

Latest revision as of 06:00, 2 November 2014

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K1321343

For reference, the cellulose binding domain binding capability of CBDclos (C-terminally fused) to bacterial cellulose was measured relative to other cellulose binding domains when fused to sfGFP, the data for which can be seen here - K1321341.

User Reviews

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