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Team:ETH Zurich/Experiments
From 2013.igem.org
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[[File:XGlcNAc_on_nagZ_t7_strain.JPG|left|200px|thumb|Fig.5: X-Gal on LacZ colony]] | [[File:XGlcNAc_on_nagZ_t7_strain.JPG|left|200px|thumb|Fig.5: X-Gal on LacZ colony]] | ||
[[File:XGlcNAc_on_nagZ_t7_strain.JPG|left|200px|thumb|Fig.5: Beta-green-X-gal on LacZ colony]] | [[File:XGlcNAc_on_nagZ_t7_strain.JPG|left|200px|thumb|Fig.5: Beta-green-X-gal on LacZ colony]] | ||
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Revision as of 11:06, 26 August 2013
Tranformation and cloning constructs
| Sender cells | Receiver cells | |
|---|---|---|
| AHL expressing system | Fluorescent reporter system | Enzyme reaction-based reporter system |
| Pcons-LuxI 1 (Strong) | Plac-LuxR-Plux-GFP | Plac-LuxR-Plux-LacZ |
| Pcons-LuxI 2 | Plac-LuxR-Plux-YFP | Plac-LuxR-Plux-Aes |
| Pcons-LuxI 3 | Plac-LuxR-Plux-CFP | Plac-LuxR-Plux-GusA |
| Pcons-LuxI 4 (Weak) | Plac-LuxR-Plux-PhoA | |
| Plac-LuxR-Plux-NagZ | ||
| Biobrick | Registry part |
|---|---|
| LuxI | K805016 |
| LuxR | C0062 |
| Pcons 1 (Strong) | J23100 |
| Pcons 2 | J23118 |
| Pcons 3 | J23110 |
| Pcons 4 (Weak) | J23114 |
| Mutagenized Plux ( low [AHL]) |
Optimized Biobrick ! |
| Mutagenized Plux ( middle [AHL]) |
Optimized Biobrick ! |
| Mutagenized Plux ( high [AHL]) |
Optimized Biobrick ! |
| pLuxR | R0062 |
| Plac | I14032 |
| GFP | E0840 |
| YFP | E0030 |
| CFP | E0020 |
| LacZ | I732006 | GusA | New Biobrick ! |
| PhoA | New Biobrick ! |
| Aes | New Biobrick ! |
| NagZ | New Biobrick ! |
Here we will paste some circuit designs
Enzyme-substrate reactions
We have cloned fluorescent receiver systems as backup for our circuit in case the hydrolase reaction do not work properly.
The enzyme substrate reactions take less than 5 minutes and are visible by eye.
Our minesweeper become better and better so keep on track for updates !
| Hydrolase | Complementary substrate / IUPAC name | Visible color | ||
|---|---|---|---|---|
| LacZ | Beta-Galactosidase | X-Gal | 5-Bromo-4chloro-3-indolyl-beta-galactopyranoside | Blue |
| LacZ | Beta-Galactosidase | Green-beta-D-Gal | N-Methyl-3-indolyl-beta-D_galactopyranoside | Green |
| GusA | Beta-glucuronidase | Magenta glucuronide | 6-chloro-3-indolyl-beta-D-glucuronide-cycloheylammonium salt | Red |
| PhoA | Alkaline phosphatase | pNPP | 4-Nitrophenylphosphatedi(tris) salt | Yellow |
| Aes | Carboxyl esterase | Magenta butyrate | 5-bromo-6-chloro-3-indoxyl butyrate | Magenta |
| NagZ | Glycoside hydrolase | X-glucosaminide X-Glunac | 5-bromo-4-chloro-3-indolyl-N-acetyl-beta-D-glucosaminide | Blue |
