Difference between revisions of "Part:BBa K523025"
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* A (possibly inactivated) cellulose binding domain | * A (possibly inactivated) cellulose binding domain | ||
* The GH6 endoglucanase domain | * The GH6 endoglucanase domain | ||
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+ | In the process of creating this part, we (Edinburgh 2011) 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: | ||
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+ | [[Image:K523025_on_congo_red.jpg|400px|center]] | ||
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+ | Evidently we had three correct colonies, here numbered 8, 17, and 22. These had already been tested for their ability to degrade MUC, in an assay similar to that for <partinfo>BBa_K523016</partinfo>. Thus, both the exoglucanase and the endoglucanase domains have been shown to work. | ||
Latest revision as of 18:52, 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 (Edinburgh 2011) 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:
Evidently we had three correct colonies, here numbered 8, 17, and 22. These had already been tested for their ability to degrade MUC, in an assay similar to that for BBa_K523016. Thus, both the exoglucanase and the endoglucanase domains have been shown to work.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 1148
Illegal NotI site found at 2846 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2759
Illegal XhoI site found at 2257
Illegal XhoI site found at 2506 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE 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 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1201
Illegal SapI.rc site found at 1284