Team:TU Delft/Project/sensing/characterization

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(New page: =Characterization= Strains: * PalkS / AlkS: E.coli K12/310C * PalkS / AlkS / PalkB: E.coli K12/311C * P(CaiF) / AlkS / PalkB: E.coli K12/312C * Negative control: ''E.coli K12'' The ...)
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=Characterization=
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__NOTOC__
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{{Team:TU_Delft/frame_check}}
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<html><center><img src="http://2010.igem.org/wiki/images/0/00/TU_Delft_project_navigation.jpg" usemap="#projectnavigation" border="0" /></center><map id="projectnavigation" name="projectnavigation"><area shape="rect" alt="Characterization" title="" coords="309,3,591,45" href="http://2010.igem.org/Team:TU_Delft#page=Project/sensing/characterization" target="" /><area shape="rect" alt="Results" title="" coords="609,3,891,44" href="http://2010.igem.org/Team:TU_Delft#page=Project/sensing/results" target="" /><area shape="rect" alt="Parts" title="" coords="9,3,290,44" href="http://2010.igem.org/Team:TU_Delft#page=Project/sensing/parts" target="" /></map></html>
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==Characterization of the Sensing Parts==
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===Characterized Strains===
Strains:  
Strains:  
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* PalkS / AlkS: E.coli K12/310C
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* P(CaiF)-E0240: E.coli K12/331A
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* PalkS / AlkS / PalkB: E.coli K12/311C
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* Negative control: ''E.coli K12'' carrying J13002 on the empty plasmid pSB1A2.
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* P(CaiF) / AlkS / PalkB: E.coli K12/312C
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* Negative control: ''E.coli K12''
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The characterization process of the hydrocarbon sensing sub-project will entail output measurements of fluorescence at varying octane concentrations (0.1% – 1%) of the above mentioned strains. These measurements will be performed using a 96-well plate reader. Growth curves will be followed by OD-600 alongside the fluorescence measurements. The obtained fluorescence and growth measurements will be analyzed and modeled accordingly.
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===[http://partsregistry.org/Part:BBa_K398331 pCaiF]===
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We implemented the pCaiF promoter to enable a metabolic switch from glucose to hydrocarbons when glucose depletion takes place in the culture. We wanted to know how was the response of this part in ''E. coli'', so we prepared an experiment in which different glucose concentrations were tested. We did that, because theoretically the glucose will be depleted at different culture times so we could check nicely the difference on GFP expression depending on the glucose depletion.
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One 96-well plate was prepared with a sextuplicate of each strain: ''E. coli'' 331D (which contains [http://partsregistry.org/Part:BBa_K398331 BBa_K398331]) and our negative control which was ''E. coli'' with J13002 in the plasmid pSB1A2. For that purpose we followed [http://2010.igem.org/Team:TU_Delft#page=Notebook/protocols&anchor=pCaiF_characterization_experiment our standard protocol].
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Go to the [http://2010.igem.org/Team:TU_Delft#page=Project/sensing/results results page] if you want to know the end of this story!!!! ;-)
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<html><center><img src="http://2010.igem.org/wiki/images/0/00/TU_Delft_project_navigation.jpg" usemap="#projectnavigation" border="0" /></center><map id="projectnavigation" name="projectnavigation"><area shape="rect" alt="Characterization" title="" coords="309,3,591,45" href="http://2010.igem.org/Team:TU_Delft#page=Project/sensing/characterization" target="" /><area shape="rect" alt="Results" title="" coords="609,3,891,44" href="http://2010.igem.org/Team:TU_Delft#page=Project/sensing/results" target="" /><area shape="rect" alt="Parts" title="" coords="9,3,290,44" href="http://2010.igem.org/Team:TU_Delft#page=Project/sensing/parts" target="" /></map></html>

Latest revision as of 16:19, 27 October 2010

CharacterizationResultsParts

Characterization of the Sensing Parts

Characterized Strains

Strains:

  • P(CaiF)-E0240: E.coli K12/331A
  • Negative control: E.coli K12 carrying J13002 on the empty plasmid pSB1A2.

pCaiF

We implemented the pCaiF promoter to enable a metabolic switch from glucose to hydrocarbons when glucose depletion takes place in the culture. We wanted to know how was the response of this part in E. coli, so we prepared an experiment in which different glucose concentrations were tested. We did that, because theoretically the glucose will be depleted at different culture times so we could check nicely the difference on GFP expression depending on the glucose depletion.

One 96-well plate was prepared with a sextuplicate of each strain: E. coli 331D (which contains BBa_K398331) and our negative control which was E. coli with J13002 in the plasmid pSB1A2. For that purpose we followed our standard protocol.

Go to the results page if you want to know the end of this story!!!! ;-)


CharacterizationResultsParts