Revision as of 20:03, 17 October 2014 by Mosthege (Talk | contribs)

X-press: Characterization of Team Heidelbergs pSBX-vectors

The iGEM team Heidelberg constructed a set of expression vectors that contain a T7 promoter prior to the BioBrick prefix. This way RBS-CDS parts can be cloned into the vectors by standard BioBrick assembly and then expressed in BL21(DE3) by induction with IPTG.

Aachen Heidelberg vector Design.png
Heidelberg X-press vector design
The iGEM Team Heidelberg created new expression vectors with an integrated T7 Promoter

In collaboration with the iGEM Team Heidelberg the iGEM Team Aachen tested 8 of these vectors for their expression capabilities.

The Team Heidelberg expression vectors

These are the tested constructs:

Plasmid Backbone Insert Length vector [bp] Length Insert [bp] Resistance
pSBX1A3-BBa_13507 pSBX1A3 (high copy) BBa_13507 3067 1226 50 mg/ml Ampicilin
pSBX1C3-BBa_13507 pSBX1C3 (high copy) BBa_13507 2982 1226 35 mg/ml Chloramphenicol
pSBX1K3-BBa_13507 pSBX1K3 (high copy) BBa_13507 3116 1226 50 mg/ml Kanamycin
pSBX1T3-BBa_13507 pSBX1T3 (high copy) BBa_13507 3322 1175 10 mg/ml Tetracyclin
pSBX4A5-BBa_13507 pSBX4A5 (low copy) BBa_13507 4307 1192 50 mg/ml Ampicilin
pSBX4C5-BBa_13507 pSBX4C5 (low copy) BBa_13507 4127 1186 15 mg/ml Chloramphenicol
pSBX4K5-BBa_13507 pSBX4K5 (low copy) BBa_13507 4331 1192 50 mg/ml Kanamycin

We also used an mRFP selection marker with a constant expression of mRFP as a positive control.

To find out more about the Results of the characterization check out the the collaboration page of Team Heidelberg.

Aachen 17-10-14 Summary iFG.PNG

Aachen 17-10-14 PSBX1A3 iFG.PNG

Aachen 17-10-14 PSBX1C3 iFG.PNG

Aachen 17-10-14 PSBX4A5 iFG.PNG

Aachen 17-10-14 PSBX4C5 iFG.PNG



Fluorescence data of pSBX cultivation