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Team:EPF-Lausanne/Our Project/Data
From 2011.igem.org
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== Other New parts == | == Other New parts == | ||
Revision as of 02:11, 22 September 2011
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 made a T7-promoter-driven lysis device by adding our wildtype T7 promoter () upstream of the Berkeley lysis cassette () inserted in the () plasmid.
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
T7 Promoter variants
Other New parts
T7 Promoter
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):
TetR mutants
In-vivo testing was used to characterise these:
- 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
Medium-strength Plac
T7 promoter variants
Pre-existing parts we improved
- Experience - The Berkeley Lysis Device, BBa_K112808: In developing our Lysis selection device, we characterised induction, and ran proof-of-principle experiments.
