Team:ETH Zurich/Experiments 7

From 2013.igem.org

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<h1>Beta-galactosidase(LacZ)</h1>
 
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<b>Chemical conversion</b><br><br>
 
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[[File:lacZhydro.png|700px|]]
 
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<b>Colorimetric response in liquid culture</b><br><br>
 
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<b>Colorimetric response on LB-Agar</b>
 
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[[File:XGAL.png|left|350px|thumb|<b>Fig.5: X-Gal on LacZ expressing colony</b>]]
 
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[[File:Green_beta_gal_on_t7_strain.JPG|left|350px|thumb|<b>Fig.6: Beta-green-X-gal on LacZ expressing colony</b>]]
 
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<h1>Alkaline phosphatase(PhoA)</h1>
 
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<b>Chemical conversion</b><br><br>
 
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[[File:phoAhydro.png|700px|]]
 
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<b>Colorimetric response in liquid culture</b><br><br>
 
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<b>Colorimetric response on LB-Agar</b>
 
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[[File:PNPP_on_phoA_t7_strain.JPG|left|350px|thumb|<b>Fig.2: NPP on PhoA expressing colony</b>]]
 
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<h1>Acetyl esterase(Aes)</h1>
 
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<b>Chemical conversion</b><br><br>
 
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[[File:Aeshydro.png|700px|]]
 
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<b>Colorimetric response in liquid culture</b><br><br>
 
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<b>Colorimetric response on LB-Agar</b>
 
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[[File:Magenta_butyrate_on_aes_t7_strain.JPG|left|324px|thumb|<b>Fig.1: Magenta butyrate on Aes expressing colony</b>]]
 
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<h1>Beta-glucuronidase(GusA)</h1>
 
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<b>Chemical conversion</b><br><br>
 
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[[File:GusAhydro.png|700px|]]
 
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<b>Colorimetric response in liquid culture</b><br><br>
 
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<b>Colorimetric response on LB-Agar</b>
 
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[[File:Salmon_glcUA_on_gusA_t7_strain.JPG |left|340px|thumb|<b>Fig.3 Magenta glucuronide on GusA expressing colony</b>]]
 
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<h1>β-N-Acetylglucosaminidase(NagZ)</h1>
 
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<b>Chemical conversion</b><br><br>
 
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[[File:nagZhydro.png|700px|]]
 
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<b>Colorimetric response in liquid culture</b><br><br>
 
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<b>Colorimetric response on LB-Agar</b>
 
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Revision as of 10:10, 16 September 2013

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What about those hydrolases ?

Colisweeper depends largely on processing of molecular signals and generation of visible and distinguishable outputs accordingly. Next to our PLuxR promoter mutations, which detect ranges of OHHL concentrations, we made use of a reporter system which gives colorimetric responses and additionally provides that the visibility of output is only to be triggered by the player of Colisweeper: A set of orthogonal hydrolases which specifically cleave the chromogenic hydrolase substrates added by the player.
These hydrolases include the Citrobacter alkaline phospohatase, the Bacillus subtilis β-glucuronidase and the Escherichia coli acetylesterase, N-acetyl-β-glucosaminidase and β-galactosidase. To ensure specific enzyme-substrate reactions, we used a triple deletion Escherichia coli strain, lacking aes (acetylesterase), gusA (β-glucuronidase) and nagZ (N-acetyl-β-glucosaminidase).