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Team:ETH Zurich/Experiments 3
From 2013.igem.org
(Difference between revisions)
| Line 14: | Line 14: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td>LacZ</td> | + | <td>[https://2013.igem.org/Team:ETH_Zurich/Experiments_4 LacZ]</td> |
<td>Beta-Galactosidase</td> | <td>Beta-Galactosidase</td> | ||
<td>X-Gal</td> | <td>X-Gal</td> | ||
| Line 21: | Line 21: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td>LacZ</td> | + | <td>[https://2013.igem.org/Team:ETH_Zurich/Experiments_4 LacZ]</td> |
<td>Beta-Galactosidase</td> | <td>Beta-Galactosidase</td> | ||
<td>Green-beta-D-Gal</td> | <td>Green-beta-D-Gal</td> | ||
| Line 28: | Line 28: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td>GusA</td> | + | <td>[https://2013.igem.org/Team:ETH_Zurich/Experiments_4 GusA]</td> |
<td>Beta-glucuronidase</td> | <td>Beta-glucuronidase</td> | ||
<td>Magenta glucuronide</td> | <td>Magenta glucuronide</td> | ||
| Line 35: | Line 35: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td>PhoA</td> | + | <td>[https://2013.igem.org/Team:ETH_Zurich/Experiments_4 PhoA]</td> |
<td>Alkaline phosphatase</td> | <td>Alkaline phosphatase</td> | ||
<td>pNPP</td> | <td>pNPP</td> | ||
| Line 42: | Line 42: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td>Aes</td> | + | <td>[https://2013.igem.org/Team:ETH_Zurich/Experiments_4 Aes]</td> |
<td>Carboxyl esterase</td> | <td>Carboxyl esterase</td> | ||
<td>Magenta butyrate</td> | <td>Magenta butyrate</td> | ||
| Line 49: | Line 49: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td>NagZ</td> | + | <td>[https://2013.igem.org/Team:ETH_Zurich/Experiments_4 NagZ]</td> |
<td>Glycoside hydrolase</td> | <td>Glycoside hydrolase</td> | ||
<td>X-glucosaminide X-Glunac</td> | <td>X-glucosaminide X-Glunac</td> | ||
Revision as of 06:18, 27 August 2013
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 | Time(s)for colorimetric response | ||
|---|---|---|---|---|---|
| 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 | |
An enzyme-substrate test matrix (Figure 6) was established to test each substrate against each enzyme. The results were as expected (Figure 6.2) and no cross reaction is visible. The NagZ-X - glucosaminide X-Glunac reveal some difficulties in the liquid culture as well as on the agar plate.
