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Team:TUDelft/MIntegration
From 2009.igem.org
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* oriTR - to allow transfer during conjugation | * oriTR - to allow transfer during conjugation | ||
* constitutively expressed trbK - to block back propagation of the signal, to reduce redundant transfer, and to prevent lethal zygosis | * constitutively expressed trbK - to block back propagation of the signal, to reduce redundant transfer, and to prevent lethal zygosis | ||
| - | * constitutively expressed trbC - if lethal zygosis turn out to be a problem | + | * ''constitutively expressed trbC - if lethal zygosis turn out to be a problem'' |
| - | * constitutively expressed GFP for the green team and RFP for the red team | + | * constitutively expressed <font color=green>GFP</font> for the green team and <font color=red>RFP</font> for the red team |
* delay device | * delay device | ||
** input: the delay is triggered by the presence of the helper plasmid | ** input: the delay is triggered by the presence of the helper plasmid | ||
** output: expression of the I-SceI homing endonuclease | ** output: expression of the I-SceI homing endonuclease | ||
* cut sites for the I-SceI homing endonuclease | * cut sites for the I-SceI homing endonuclease | ||
| + | * a low copy backbone to allow the endonuclease to destroy the signal plasmid | ||
==Helper Plasmid== | ==Helper Plasmid== | ||
| - | This will consist of a modified R751 plasmid | + | This will consist of a modified R751 plasmid with the following features: |
| - | + | * trbK knockout to allow cells containing the helper plasmid to conjugate | |
| + | * oriTR knockout to prevent transfer of the helper plasmid during conjugation | ||
| + | * ''trbC knockout to prevent lethal zygosis'' | ||
| + | * constitutively expressed trigger for the delay device | ||
[[Team:TUDelft | <<< Back Home]] | [[Team:TUDelft | <<< Back Home]] | ||
{{Template:TUDelftiGEM2009_end}} | {{Template:TUDelftiGEM2009_end}} | ||
Revision as of 21:52, 19 October 2009
Module Integration
In order for us to achieve our goal of having a relay race between GFP and RFP teams,
various parts from the the three modules must be integrated together into a single working system. What follows is a description of the requirements for the integrated system.
Signal Plasmid
The signal plasmid will contain the following features:
- oriTR - to allow transfer during conjugation
- constitutively expressed trbK - to block back propagation of the signal, to reduce redundant transfer, and to prevent lethal zygosis
- constitutively expressed trbC - if lethal zygosis turn out to be a problem
- constitutively expressed GFP for the green team and RFP for the red team
- delay device
- input: the delay is triggered by the presence of the helper plasmid
- output: expression of the I-SceI homing endonuclease
- cut sites for the I-SceI homing endonuclease
- a low copy backbone to allow the endonuclease to destroy the signal plasmid
Helper Plasmid
This will consist of a modified R751 plasmid with the following features:
- trbK knockout to allow cells containing the helper plasmid to conjugate
- oriTR knockout to prevent transfer of the helper plasmid during conjugation
- trbC knockout to prevent lethal zygosis
- constitutively expressed trigger for the delay device
