"
Team:EPF-Lausanne/Our Project/Data
From 2011.igem.org
Data
team, please check this page: [1]
Contents |
Overview
Summary
There are three pillars that underlie our transcription factor development pipeline. The first pillar is a selection system which lyses cells containing strong matches between a transcription factor and its promoter and allows for the speedy recovery of the relevant DNA. The second pillar is an in vivo characterization system that uses two different reporter systems as a way to document the binding affinities of the TetR transcription factor and its promoter pTet. The third pillar is an in-vitro characterization system called MITOMI that allows for high-throughput analysis of DNA-protein interaction strengths.
In developing these three pillars, a number of parts were made. We have listed all of them and have highlighted our favorites.
Systems
Selection System: We cloned the Berkeley Lysis cassette (BBa_K112808) under control of the T7 promoter into a low copy number plasmid (pSB3K1, formerly BBa_I739202).
In Vivo Characterization:
In Vitro Characterization:
Biobricks
Favourite parts
Pre-existing parts characterised
- Experience - The Berkeley Lysis Device, BBa_K112808: In developing our Lysis selection device, we characterised induction, and ran proof-of-principle experiments.
All other parts submitted
TetR Mutants
- V36F mutant: K613013
- V36F W43S double mutant: K613014
- E37A W43S T141A triple mutant: K613015
- P39K mutant: K613016
- Y42F K108E double mutant: K613018
- P39Q Y42M double mutant: K613019
T7 Promoter variants
Our lysis selection device uses what we refer to as the wild-type T7 Promoter:
In addition, we submitted the following mutants:
Variants without additional lac operator (constitutive):
Variants with additional lac operator (LacI repressed):
Medium-strength Plac
