Difference between revisions of "Part:BBa K1321356:Experience"
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Edinburgh 2015 iGEM team | Edinburgh 2015 iGEM team | ||
| − | We improved and expanded characterisation on the part BBa_K1321356, a cellulose binding domain (CBD) submitted by the Imperial College 2014 team. After confirming expression through SDS PAGE we incubated iso-standard paper chads from Whatman 54 filter paper in a crude cell lysate containing the CBD bound to super folded GFP for ten minutes. We then let the protein saturated chads sit in a large volume of 1 x PBS buffer at pH 7.4 (to remain consistent with Imperial’s initial characterisation.) At different time points 5 chads were taken from the buffer and placed into a 96 well plate, over 30 minutes. The decrease in fluorescence over time was indicative of relative binding affinity, to Whatman 54 paper. The graph below shows our results. | + | <br>We improved and expanded characterisation on the part BBa_K1321356, a cellulose binding domain (CBD) submitted by the Imperial College 2014 team. After confirming expression through SDS PAGE we incubated iso-standard paper chads from Whatman 54 filter paper in a crude cell lysate containing the CBD bound to super folded GFP for ten minutes. We then let the protein saturated chads sit in a large volume of 1 x PBS buffer at pH 7.4 (to remain consistent with Imperial’s initial characterisation.) At different time points 5 chads were taken from the buffer and placed into a 96 well plate, over 30 minutes. The decrease in fluorescence over time was indicative of relative binding affinity, to Whatman 54 paper. The graph below shows our results. |
Latest revision as of 08:50, 22 September 2015
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Applications of BBa_K1321356
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 (here) - K1321346.
Edinburgh 2015 iGEM team
We improved and expanded characterisation on the part BBa_K1321356, a cellulose binding domain (CBD) submitted by the Imperial College 2014 team. After confirming expression through SDS PAGE we incubated iso-standard paper chads from Whatman 54 filter paper in a crude cell lysate containing the CBD bound to super folded GFP for ten minutes. We then let the protein saturated chads sit in a large volume of 1 x PBS buffer at pH 7.4 (to remain consistent with Imperial’s initial characterisation.) At different time points 5 chads were taken from the buffer and placed into a 96 well plate, over 30 minutes. The decrease in fluorescence over time was indicative of relative binding affinity, to Whatman 54 paper. The graph below shows our results.
User Reviews
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