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Team:EPF-Lausanne/Our Project/Data
From 2011.igem.org
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== Biobricks == | == Biobricks == | ||
| - | + | == Favourite parts == | |
[http://partsregistry.org/Part:BBa_K613015 BBa_K613015] | [http://partsregistry.org/Part:BBa_K613015 BBa_K613015] | ||
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[http://partsregistry.org/Part:BBa_K613017 BBa_K613017] | [http://partsregistry.org/Part:BBa_K613017 BBa_K613017] | ||
| - | + | == Pre-existing parts characterised == | |
* [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. | * [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. | ||
| - | + | == All other parts submitted == | |
| - | + | === TetR Mutants === | |
* V36F mutant: [http://partsregistry.org/Part:BBa_K613013 K613013] | * V36F mutant: [http://partsregistry.org/Part:BBa_K613013 K613013] | ||
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* P39Q Y42M double mutant: [http://partsregistry.org/Part:BBa_K613019 K613019] | * P39Q Y42M double mutant: [http://partsregistry.org/Part:BBa_K613019 K613019] | ||
| - | + | === T7 Promoter variants === | |
Our lysis selection device uses what we refer to as the wild-type T7 Promoter: | Our lysis selection device uses what we refer to as the wild-type T7 Promoter: | ||
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[http://partsregistry.org/Part:BBa_K613012 K613012] | [http://partsregistry.org/Part:BBa_K613012 K613012] | ||
| - | + | === Medium-strength Plac === | |
[http://partsregistry.org/Part:BBa_K613000 K613000] | [http://partsregistry.org/Part:BBa_K613000 K613000] | ||
Revision as of 02:16, 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 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
[http://partsregistry.org/Part:BBa_K613015 BBa_K613015]
[http://partsregistry.org/Part:BBa_K613016 BBa_K613016]
[http://partsregistry.org/Part:BBa_K613017 BBa_K613017]
Pre-existing parts characterised
- [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.
All other parts submitted
TetR Mutants
- V36F mutant: [http://partsregistry.org/Part:BBa_K613013 K613013]
- V36F W43S double mutant: [http://partsregistry.org/Part:BBa_K613014 K613014]
- Y42F K108E double mutant: [http://partsregistry.org/Part:BBa_K613018 K613018]
- P39Q Y42M double mutant: [http://partsregistry.org/Part:BBa_K613019 K613019]
T7 Promoter variants
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]
Medium-strength Plac
[http://partsregistry.org/Part:BBa_K613000 K613000]
