Difference between revisions of "Part:BBa K523025"

Revision as of 18:33, 28 October 2011

Plac + RBS + cxnA (cex + cenA fusion)

Lac promoter and RBS, followed by Cex exoglucanase from Cellulomonas fimi fused to CenA endoglucanase from the same species. The coding sequence contains, in order:

The GH10 exoglucanase domain
A (possibly inactivated) cellulose binding domain
The GH6 endoglucanase domain

In the process of creating this part, we tested putative correct clones on carboxymethylcellulose (CMC) plates, with Congo red dye. This test produces a zone of clearing if CMC is degraded by endoglucanase. We had three successful colonies:

Evidently we had three correct colonies (these had already been tested for their ability to degrade MUC, in an assay similar to that for BBa_K523016.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NotI site found at 1148
Illegal NotI site found at 2846
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 2759
Illegal XhoI site found at 2257
Illegal XhoI site found at 2506
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 781
Illegal NgoMIV site found at 1154
Illegal NgoMIV site found at 1656
Illegal NgoMIV site found at 2037
Illegal NgoMIV site found at 2962
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 1201
Illegal SapI.rc site found at 1284

@@ Line 8: / Line 8: @@
 * The GH6 endoglucanase domain
+In the process of creating this part, we tested putative correct clones on carboxymethylcellulose (CMC) plates, with Congo red dye. This test produces a zone of clearing if CMC is degraded by endoglucanase. We had three successful colonies:
+[[File:K523025_on_congo_red.jpg|400px|center]]
+Evidently we had three correct colonies (these had already been tested for their ability to degrade MUC, in an assay similar to that for <partinfo>BBa_K523016</partinfo>.
 <!-- Add more about the biology of this part here