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Team:EPF-Lausanne/Our Project/Data
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=== Systems === | === Systems === | ||
| - | Selection System: | + | 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 Vivo Characterization: | ||
Revision as of 02:10, 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:
New Favourite Parts
These are officially our three favourite parts:
[http://partsregistry.org/Part:BBa_K613015 BBa_K613015]
[http://partsregistry.org/Part:BBa_K613016 BBa_K613016]
[http://partsregistry.org/Part:BBa_K613017 BBa_K613017]
Other New parts
T7 Promoter
Our lysis selection device uses what we refer to as the wild-type T7 Promoter:
[http://partsregistry.org/Part:BBa_K613001 K613001]
In addition, we submitted the following mutants:
Variants without additional lac operator (constitutive):
[http://partsregistry.org/Part:BBa_K613002 K613002]
[http://partsregistry.org/Part:BBa_K613003 K613003]
[http://partsregistry.org/Part:BBa_K613004 K613004]
[http://partsregistry.org/Part:BBa_K613005 K613005]
[http://partsregistry.org/Part:BBa_K613006 K613006]
Variants with additional lac operator (LacI repressed):
[http://partsregistry.org/Part:BBa_K613007 K613007]
[http://partsregistry.org/Part:BBa_K613008 K613008]
[http://partsregistry.org/Part:BBa_K613009 K613009]
[http://partsregistry.org/Part:BBa_K613010 K613010]
[http://partsregistry.org/Part:BBa_K613011 K613011]
[http://partsregistry.org/Part:BBa_K613012 K613012]
TetR mutants
In-vivo testing was used to characterise these:
- V36F mutant: [http://partsregistry.org/Part:BBa_K613013 K613013]
- V36F W43S double mutant: [http://partsregistry.org/Part:BBa_K613014 K613014]
- E37A W43S T141A triple mutant: [http://partsregistry.org/Part:BBa_K613015 K613015]
- P39K mutant: [http://partsregistry.org/Part:BBa_K613016 K613016]
- Y42F K108E double mutant: [http://partsregistry.org/Part:BBa_K613018 K613018]
- P39Q Y42M double mutant: [http://partsregistry.org/Part:BBa_K613019 K613019]
Medium-strength Plac
[http://partsregistry.org/Part:BBa_K613000 K613000]
T7 promoter variants
Pre-existing parts we improved
- [http://partsregistry.org/Part:BBa_K112808:Experience Experience] - The Berkeley Lysis Device, BBa_K112808: In developing our Lysis selection device, we characterised induction, and ran proof-of-principle experiments.
