Difference between revisions of "Part:BBa K1321356:Experience"

(→‎Applications of BBa_K1321356)
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
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
Line 6: Line 5:
 
===Applications of BBa_K1321356===
 
===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 ([https://parts.igem.org/Part:BBa_K1321348 here])  - K1321346.
 
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 ([https://parts.igem.org/Part:BBa_K1321348 here])  - K1321346.
 +
 +
 +
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.
 +
 +
 +
[[File:5.20K.jpg]]
  
 
===User Reviews===
 
===User Reviews===

Revision as of 01:18, 22 September 2015

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_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.


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.


5.20K.jpg

User Reviews

UNIQ225d37a4aba1a7ea-partinfo-00000000-QINU UNIQ225d37a4aba1a7ea-partinfo-00000001-QINU