"
Team:Tuebingen/Project/Plasmids
From 2013.igem.org
We have designed our system to consist of three plasmids that will be be transformed into S. cerevisiae. Each plasmid will contain one part of our project - receptor, inverter, reporter - whereby there will be two receptor plasmids (one with mPR Dr, one with mPR Xl), two inverter plasmids (one with ROX1, one with MIG1), and two reporter plasmids (one with Psuc2, one with Panb1).
All our constructs are cloned into the pRS plasmid. The pRS plasmid is derived from the E. coli plasmid pBLUESCRIPT (Stratagene, La Jolla, CA, USA) and serves as a shuttle vector between Saccharomyces cerevisiae and bacteria like E. coli (Sikorski and Hieter, 1989). pRS offers built in blue/white screening and is a high copy plasmid in bacteria (Sikorski and Hieter, 1989) thus plasmid extractions from E. coli yield high quantities of plasmid DNA that can be used for yeast transformations.
Each plasmid will carry three parts, namely a promoter, a protein coding sequence, and the ADH1 terminator (Tadh1). We have retreived Tadh1 from the Registry - BBa_K801012.
"Receptor"-Plasmids
pRS313 containing Padh1, mPR Dr, and Tadh1:
pRS313 containing Padh1, mPR Xl, and Tadh1:
"Inverter"-Plasmids
pRS315 containing Pfet3, rox1, and Tadh1:
pRS315 containing Pfet3, mig1, and Tadh1:
"Reporter"-Plasmids
pRS316 containing Panb1, luciferase (luc), and Tadh1:
pRS316 containing Psuc2, luciferase (luc), and Tadh1:
References
SIKORSKI, R. S. & HIETER, P. 1989. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics, 122, 19-27.
Back to top
