Team:ETH Zurich/Experiments 7

From 2013.igem.org

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<h1>β-Glucuronidase (GusA)</h1>
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<p align="justify"></p> β-Glucuronidase (gusA) from Bacillis Subtilis
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gusA (also called uidA[1]) encodes β-Glucuronidase, an intracellular enzyme that catalyzes the hydrolysis of β-D-glucuronides.
<h1>Alkaline phosphatase (PhoA)</h1>
<h1>Alkaline phosphatase (PhoA)</h1>
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<p align="justify"></p>Alkaline Phosphatase (phoA) from Citrobacter
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<h1>β-Glucuronidase (GusA)</h1>
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The alkaline phosphatase is a periplasmic homodimeric hydrolase. Each monomer contains 429 amino acids.
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<h1>Acetylesterase (Aes)</h1>
<h1>Acetylesterase (Aes)</h1>
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<h1>β-N-Acetylglucosaminidase (NagZ)</h1>
<h1>β-N-Acetylglucosaminidase (NagZ)</h1>
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<p align="justify"></p> beta-N-Acetylglucosaminidase (nagZ) from Escherichia Coli
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nagZ encodes β-N-Acetylglucosaminidase, a cytoplasmatic hydrolase which is involved in the murein tripeptide recycling pathway of E.coli
<h1>β-Galactosidase (LacZ)</h1>
<h1>β-Galactosidase (LacZ)</h1>
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Revision as of 18:09, 17 September 2013

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Contents

What about those hydrolases ?

Colisweeper depends largely on processing of molecular signals and generation of visible and distinguishable color outputs used as logic to play the game. Next to our PLuxR promoter mutations, which detect ranges of OHHL concentrations, we make use of a reporter system which gives colorimetric responses only to be triggered by the player. As our reporter system, we rely on a set of orthogonal hydrolase enzymes that are native to Escherichia coli. These hydrolases include the Citrobacter alkaline phospohatase, the Bacillus subtilis β-glucuronidase and the Escherichia coli acetylesterase, β-N-Acetylglucosaminidase and β-galactosidase. In order to prevent background expression of these native hydrolases in Escherchia coli, we use a triple knockout strain that has three hydrolase genes knocked out: gusA (β-glucuronidase), aes (acetylesterase) and nagZ (β-N-acetylglucosaminidase). Addition of a multi-substrate mix by the player leads to an enzyme-susbtrate reaction which specifically cleaves the chromogenic substrates, thereby producing a visible color output.


β-Glucuronidase (GusA)

β-Glucuronidase (gusA) from Bacillis Subtilis

gusA (also called uidA[1]) encodes β-Glucuronidase, an intracellular enzyme that catalyzes the hydrolysis of β-D-glucuronides.

Alkaline phosphatase (PhoA)

Alkaline Phosphatase (phoA) from Citrobacter

The alkaline phosphatase is a periplasmic homodimeric hydrolase. Each monomer contains 429 amino acids.

Acetylesterase (Aes)

β-N-Acetylglucosaminidase (NagZ)

beta-N-Acetylglucosaminidase (nagZ) from Escherichia Coli

nagZ encodes β-N-Acetylglucosaminidase, a cytoplasmatic hydrolase which is involved in the murein tripeptide recycling pathway of E.coli

β-Galactosidase (LacZ)


References