Revision as of 22:52, 27 October 2010

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.

Characterization of BBa_K374006

Positive feedback antitermination on double reporter plasmid

The iGEM DTU team 2010 have submitted this part, after using it in the development of a genetic bistable switch.

Aim and strategy

The aim of the characterization was to asses if this part together with BBa_K374005 could have an antiterminating function that could be used as a regulatory mechanism. We characterized the functionality of the part in the following construct.

The test plasmid was constructed to biobrick standards, for more information see our wiki, GFP and RFP was used as reporter proteins. The distance from the boxB site in the nut site (marked with a grey triangle) to the terminator was 26bp. The RBS sites was the BB standard RBS BBa_B0034, except for the N protein where the natural RBS site was used. 5 constructs where produced named from A to E, with three different terminator strengths and references with out terminator and N protein.

we tested if:

If it was possible to trigger a positive feedback mechanism of N protein, by varying the promoter strength with the synthetic promoter library (SPL) and thus antitermination and constant expression of the down stream RFP.
If it was possible to trigger this positive feed back mechanism by inducing the N protein from a second plasmid insert with the pBAD promoter and the N protein.

Results

Conclusionc and Recommendations

User Reviews

UNIQfadb7fc9c3aa3d2d-partinfo-00000001-QINU

BBa_K374006 1 Not understood iGEM DTU 2010

We have characterized this part.

;

UNIQfadb7fc9c3aa3d2d-partinfo-00000003-QINU

@@ Line 10: / Line 10: @@
 ==== Aim and strategy ====
-The aim of the characterization was to asses if this part together with <partinfo>BBa_K374005 </partinfo> could have an antiterminating function that could be used as a regulatory mechanism. We characterized the functionality of the part in the following construct. [[Image:AntiT1.png|center|thumb|400px|Caption test to sese the text]]
+The aim of the characterization was to asses if this part together with <partinfo>BBa_K374005 </partinfo> could have an antiterminating function that could be used as a regulatory mechanism. We characterized the functionality of the part in the following construct. [[Image:AntiT1.png|center|thumb|400px|The test plasmid was constructed to biobrick standards, for more information see our wiki, GFP and RFP was used as reporter proteins. The distance from the boxB site in the nut site (marked with a grey triangle) to the terminator was 26bp. The RBS sites was the BB standard RBS BBa_B0034, except for the N protein where the natural RBS site was used. 5 constructs where produced named from A to E, with three different terminator strengths and references with out terminator and N protein.]]
-The test plasmid was constructed to biobrick standards, for more information see our wiki, GFP and RFP was used as reporter proteins. The distance from the boxB site in the nut site to the terminator was 26bp. The RBS sites was the BB standard RBS BBa_B0034, except for the N protein where the natural RBS site was used. 5 constructs where produced named from A to E, with three different terminator strengths and references with out terminator and N protein.
 we tested if: