Difference between revisions of "Part:BBa K1615098"

Latest revision as of 00:54, 22 September 2015

mRFP fused to CBDclos in RFC25

mRFP (BBa_K1615089) fused to the N terminal of CBDclos (BBa_K1321002). If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.

To test the RFP folding when fused to a CBD we placed 200µL of crude cell lysate in a 96 well plate and observed the fluorescence under a blue light transilluminator.

We tested the dissociation of this part on Whatman 54 using ISO standard chads. This involved incubating the chads in the crude cell lysate for 10 minutes and then washing them with PBS for various periods time to test the binding affinity of this CBD. The results show a very low binding affinity of CBDclos.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K1615098 short</partinfo>
-mRFP fused N terminally to CBDclos.
+mRFP (BBa_K1615089) fused to the N terminal of CBDclos (BBa_K1321002). If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.
+<br>
+<br>
+[[File:CBD RFP folding.jpg]]
+<br>
+<br>
+To test the RFP folding when fused to a CBD we placed 200µL of crude cell lysate in a 96 well plate and observed the fluorescence under a blue light transilluminator.
+<br>
+<br>
+[[File:BBa_K1615098.jpg]]
+<br>
+<br>
+We tested the dissociation of this part on Whatman 54 using ISO standard chads. This involved incubating the chads in the crude cell lysate for 10 minutes and then washing them with PBS for various periods time to test the binding affinity of this CBD. The results show a very low binding affinity of CBDclos.
 <!-- Add more about the biology of this part here