Team:ETH Zurich/Experiments
From 2013.igem.org
Cloned constructs
For the final Colisweeper circuit we plan a four plasmid system. The mine cells constitutively express LuxI and NagZ. In the Non-Mine cells LuxR and PhoA are expressed constitutively whereas Aes and GusA are expressed from pLux promoters with different sensitivities. To get to this system we tested different versions of the circuit with GFP. In the following table we list all the biobricks we used, the plasmids we cloned and what experiments we used them for. In general we used standard biobrick cloning techniques which you can find in the Methods section. Whenever we used PCR gene amplification for cloning, we list the primers used in the following table. To be able to co-transform different plasmids we used backbones with compatible origins of replication and resistance genes. In the table you can find which versions we used for which constructs.
LuxI generating constructs | |||
---|---|---|---|
Description | Cloning | Maps | |
4 | Sender cell construct with strongest constitutive promoter for GFP and Hydrolase experiments | BBa_J23100 backbone (SpeI, PstI) and BBa_K805016 insert (XbaI, PstI) | ... |
5 | Sender cell construct with second strongest constitutive promoter for GFP and Hydrolase experiments | BBa_J23118 backbone (SpeI, PstI) and BBa_K805016 insert (XbaI, PstI) | ... |
6 | Sender cell construct with second weakest constitutive promoter for GFP and Hydrolase experiments | BBa_J23110 backbone (SpeI, PstI) and BBa_K805016 insert (XbaI, PstI) | ... |
7 | Sender cell construct with weakest constitutive promoter for GFP and Hydrolase experiments | BBa_J23114 backbone (SpeI, PstI) and BBa_K805016 insert (XbaI, PstI) | ... |