Team:BOKU-Vienna/Composite Part
Composite Part
Mycolactone Riboswitch Composite Parts
To test the properties of our transcriptional Mycolactone Riboswitch BBa_K3015000 we, of course, had to incooperate them
into an expression cassette, meaning adding a Promoter to the 3’ end and a Ribosome Binding Site,
coding sequence and Terminator to the 5’ end.
We designed the composite Part with a Constituitive Promotor, the Mycolactone Riboswitch and the
Ribosome Binding Site; in that way the expression of any downstream located gene can be controlled
by the presence of Mycolactone
BBa_K3015001
We conducted the messurement, to characterize our Riboswitch with
BBa_K3015002
fused to GFP und a Terminator.
| Part number | Part name |
|---|---|
| BBa_K3015001 | pMyco-RBS |
| BBa_K3015002 | pMyco-RBS-GFP-Term |
Theophylline Riboswitch Composite Parts
To prove the functionality of our system, we also conducted experiments with a transcriptional
Theophylline-Riboswitch. [1]
We added the same Promoter and RBS down as to the Mycolactone-Riboswitch mentioned above,
creating
BBa_K3015005.
We also decided to messsure the probability of this Part by fusing it to GFP BBa_K3015007
.
Additionally, we fused it to the coding sequence of a T7-Polymerase, to induce expression of
genes following a T7-Polymerase-dependent promotor, creating
BBa_K3015008
.
| Part number | Part name |
|---|---|
| BBa_K3015005 | pTheo-RBS |
| BBa_K3015007 | pTheo-RBS-GFP-Term |
| BBa_K3015008 | pTheo-RBS-T7-Term |
T7-Promoter-amilCP composite Part for a visual readout BBa_K3015003
This composite Part BBa_K3015003 expresses the blue chromoprotein amilCP in the presence of
T7-Polymerase.
It plays an important role in our system, since this is the Part that allows the visuall readout, if
the toxin is present.
| Part number | Part name |
|---|---|
| BBa_K3015003 | T7prom-amilCP-Term |
Riboswitch Sensor cassette BBa_K3015011
This is the genetic device we realized in the lab with a transcriptional Theophylline-Riboswitch. T7-Polymerase will be expressed in the Presence of Theophylline, which will further induce the strong expression of amilCP, a blue chromoprotein, that acts as visual readout. The designed and constructed system can easily be adapted for detection of various disease-causing agents by just changing the riboswitch-sequence according to the target.
| Part number | Part name |
|---|---|
| BBa_K3015011 | Riboswitch Sensor cassette |
References:
[1] Manja Wachsmuth, et al. (2016) "Design criteria for synthetic riboswitches acting on transcription" doi: 10.1080/15476286.2015.1017235
