Composite

Part:BBa_K1321357:Experience

Designed by: Chris N Micklem   Group: iGEM14_Imperial   (2014-10-08)
Revision as of 19:56, 7 November 2015 by LiusaidhOwen (Talk | contribs)

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Applications of BBa_K1321357

Edinburgh 2015 iGEM team
We improved and expanded characterisation on the part BBa_K1321357, 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 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.
Dissociation curve of sfGFP-CBDcex.jpg



User Reviews

UNIQ739502ba48aa50a0-partinfo-00000000-QINU UNIQ739502ba48aa50a0-partinfo-00000001-QINU