Difference between revisions of "Part:BBa K1321346:Experience"

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===Applications of BBa_K1321346===
 
===Applications of BBa_K1321346===
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As part of our project we carried out an assay to determine the relative binding ability of CBD-sfGFP fusions to bacterial cellulose.   
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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.   
  
 
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])   
 
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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It was determined that the fusion with CBDclos was, on average, our fourth strongest CBD, in comparison other CBDs tested, also fused to sfGFP.
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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 fifth strongest CBD, in comparison other CBDs tested.  
  
  
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[[File:IC14_-_dH2Obplot1.png|600px|thumb|left|Figure 1 - CBD-sfGFP binding strength after three dH2O washes]]
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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-sfGFP binding strength after three 70%Ethanol 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-sfGFP binding strength after three PBS 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|left|Figure 4 - CBD-sfGFP binding strength after three BSA washes]]
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[[File:IC14-BSAbplot1.png|600px|thumb|left|Figure 4 - CBD binding strength after three BSA washes]]
  
 
===User Reviews===
 
===User Reviews===

Latest revision as of 05:47, 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_K1321346

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.

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 N-terminal CBDclos was, on average, our fifth strongest CBD, in comparison other CBDs tested.


Figure 1 - CBD binding strength after three dH2O washes.
Figure 2 - CBD binding strength after three 70%Ethanol washes
Figure 3 - CBD binding strength after three PBS washes
Figure 4 - CBD binding strength after three BSA washes

User Reviews

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