Imperial/Infector Detector/Design

From 2007.igem.org

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|Lifespan
|Lifespan
|System must have a shelf life of 7 days
|System must have a shelf life of 7 days
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|Protease Inhibitor of Cell Extract Should ensure degradation of Visual Reporter is Minimal<br> Proper Packaging should ensure that evaporation of Cell Free system  is so low that <br> system can surive for 7 days
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|Protease Inhibitor of Cell Extract should ensure degradation of Visual Reporter is Minimal<br> Proper Packaging should ensure that evaporation of Cell Free system  is so low that system can surive for 7 days
|Chassis
|Chassis
|- style="background:#eeeeff"
|- style="background:#eeeeff"
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|Sensitive AHL receiver to detect low AHL levels
|Sensitive AHL receiver to detect low AHL levels
|Construct
|Construct
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-
 
|- style="background:#eeeeff"
|- style="background:#eeeeff"
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|
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|Outputs
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|
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|System must give a visual signal if bacteria is present
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|Couple AHL to a reporter system expressing fluoresent protein eg. RFP
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|Construct
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|- style="background:#eeeeff"
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|Operating Conditions
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|System must operate within temperature 20-30°C
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|Gene expression systems and protein must be thermostable
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|Construct
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|- style="background:#eeeeff"
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|
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|System must operate within pH range of 6-8
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|Gene expression systems and protein must be pH resistant
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|Construct
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|- style="background:#eeeeff"
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|Response Time
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|System needs to have a response time under 1 hour
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|To be Determined - this is hard to design for
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|Both
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|}
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== DNA Constructs ==
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{|border=1 width=90% align="center"
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|-  style="background:#eeeeff"
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|width="400px"|'''Construct Design'''
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|width="300px"|'''Availability'''
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|-
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|width="30%"|<center>[[Image:Imperial CBD Design CBD1.png ]]</center>
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|width="--"|This part can be found as [http://partsregistry.org/Part:BBa_I13522 BBa_I13522] in the registry.
|}
|}
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While
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== Gene Construct ==
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==Chassis Selection==
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{| style="width:800px;" cellpadding="2" cellspacing="0" align="center"
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|- style="background:#ffffff; text-align:left;" color:white;" align="center"
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|| [[Image:Icgems-bulksol.png|180px]]
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||
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We have chosen to use the commerciall available S30 ''E. coli'' cell extract made by Promega.  After having looking into a variety of different [[Imperial/Cell-Free/Whatis|cell-free chassis]], we feel that this chassis best suits our needs.  In particular this chassis allows us to meet our base requirement of complying with the Health and Safety regulations of the field we are working in, as we do not want replicative bacteria that could potentially be pathogenic to come in contact with urinary catheters.
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|- style="background:#ffffff; text-align:left;" color:white;" align="center"
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|style="font-size:0.85em;"| Commercial S30 ''E. coli'' Cell Extract in bulk solution + packaging to last 7 days
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||
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In addition to complying with health regulations the S30 cell extract is commercially available meaning that it has been shown to work.  This is very important for us as it allows our focus to be on tuning the chassis to suit our needs rather than making the chassis work in the first place.
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|}
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<br clear="all">
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== Chassis Selection ==
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== References ==

Revision as of 22:36, 21 October 2007


Infector Detector: Design

Design Overview

InfectorDetectorDesignWiki.png


Property Value Design Solution System Level
Health Regulations System Must not be living replicating bacteria Use a Cell Free System e.g. Promega's S30 Cell Extract Chassis
Lifespan System must have a shelf life of 7 days Protease Inhibitor of Cell Extract should ensure degradation of Visual Reporter is Minimal
Proper Packaging should ensure that evaporation of Cell Free system is so low that system can surive for 7 days
Chassis
Inputs System must be sensitive to AHL concentration between 5-50nM Sensitive AHL receiver to detect low AHL levels Construct
Outputs System must give a visual signal if bacteria is present Couple AHL to a reporter system expressing fluoresent protein eg. RFP Construct
Operating Conditions System must operate within temperature 20-30°C Gene expression systems and protein must be thermostable Construct
System must operate within pH range of 6-8 Gene expression systems and protein must be pH resistant Construct
Response Time System needs to have a response time under 1 hour To be Determined - this is hard to design for Both



DNA Constructs

Construct Design Availability
Imperial CBD Design CBD1.png
This part can be found as BBa_I13522 in the registry.


While


Chassis Selection

Icgems-bulksol.png

We have chosen to use the commerciall available S30 E. coli cell extract made by Promega. After having looking into a variety of different cell-free chassis, we feel that this chassis best suits our needs. In particular this chassis allows us to meet our base requirement of complying with the Health and Safety regulations of the field we are working in, as we do not want replicative bacteria that could potentially be pathogenic to come in contact with urinary catheters.

Commercial S30 E. coli Cell Extract in bulk solution + packaging to last 7 days

In addition to complying with health regulations the S30 cell extract is commercially available meaning that it has been shown to work. This is very important for us as it allows our focus to be on tuning the chassis to suit our needs rather than making the chassis work in the first place.



References