Team:BCCS-Bristol/Notebook/Biobricks

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Revision as of 01:00, 20 October 2009

BCCS-Bristol
iGEM 2009

Parts Submitted to The Registry

Finished biobricks to be submitted to the iGEM parstregistry at the competition are outlined in the tables below :

The table illustrates the finished biobricks that were created as a proof of principle to illustrate OMV directed delivery.
Finished Assemblies
AraC-RBS-FhuA-GFP-Terminator
AraC-RBS-FhuA-GFP

The table illustrates biobricks that were created in the duration of the project and that will be of use to synthetic biologists.
Promoter-RBS	Promoter-RBS-Carriers	Reporters	Bioscaffolds	Proteins
AraC-RBS	AraC-RBS-OsmE	GFP-Terminator	BpuEI-BseRI	FhuA
-	AraC-RBS-FhuA	-	BpuEI-CspCI(3 versions)	OsmE

Why these biobricks?

We choose the AraC promoter because its activity can be controlled by the Arabinose levels (direct relationship)just like o rheostat. This means that we will be able to synthesize our fusion protein at will (i.e. when bacteria cell numbers are high) and at the required levels that won't cause cell lysis or non-specific aggregation in cytoplasm. The RBS J61100 is a ribosome binding site with strong affinity for RNA polymerase ensuring that once RNA polymerase binds to the RBS it will stay bound and won;t dissociate giving us maximum transcription. The Bioscaffold BBa_K259002 is used for the efficient joining of the carrier and cargo partners in the protein fusion leaving a scarless region and a flexible Gly-Ser-Gly (http://partsregistry.org/Part:BBa_K259002 more information). Also BBa_B0014 double terminator is used to stop transcription ensuring that no other DNA sequences are incorporated into our fusion protein that may interfere with the folding of the individual proteins. - pSB1A2, pSB1A3, pSB2K3 (vectors for fusion protein)

Stucture of our Biobrick

Assembly Plan (timeline)

KEY: V=vector, I=insert

@@ Line 60: / Line 60: @@
+===Why these biobricks?===
+We choose the AraC promoter because its activity can be controlled by the Arabinose levels (direct relationship)just like o rheostat. This means that we will be able to synthesize our fusion protein at will (i.e. when bacteria cell numbers are high) and at the required levels that won't cause cell lysis or non-specific aggregation in cytoplasm. The RBS J61100 is a ribosome binding site with strong affinity for RNA polymerase ensuring that once RNA polymerase binds to the RBS it will stay bound and won;t dissociate giving us maximum transcription. The Bioscaffold BBa_K259002 is used for the efficient joining of the carrier and cargo partners in the protein fusion leaving a scarless region and a flexible Gly-Ser-Gly ([[http://partsregistry.org/Part:BBa_K259002 more information]]). Also BBa_B0014 double terminator is used to stop transcription ensuring that no other DNA sequences are incorporated into our fusion protein that may interfere with the folding of the individual proteins.
+- pSB1A2, pSB1A3, pSB2K3 (vectors for fusion protein)
 ==Stucture of our Biobrick==