Team:Goettingen/Notebook/reporterSystems October
09.10.18
In the morning: Inoculation of 4 ml LB with BP270 containing pAC7::PyciAB-xylR and BP270 containing the empty BP270.
In the evening: Inoculation of 4 ml CS-Glc medium with 100 µL of the first culture from the morning. Incubation: shaking over night at 37°C.
10.10.18
Measurment of the OD600 of the over night culutures.
| Strain | OD600 |
|---|---|
| BP270 + pAC7I | 1.48 |
| BP270 + pAC7::PyciAB-xylR | 1.45 |
10 ml C minimal medium containing different glyphosate concentrations (0mM, 0.5 mM and 1 mM) were inoculated with the two strains in a 100 ml shake flasks to an OD600 of about 0.1. They were grown until they reached an OD600 of 0.5-0.8
| Strain | OD600 |
|---|---|
| BP270 + pAC7 0 mM glyphosate | 0.844 |
| BP270 + pAC7 0.5 mM glyphosate | 0.748 |
| BP270 + pAC7 1 mM glyphosate | 0.58 |
| BP270 + pAC7::PyciAB-xylR 0 mM glyphosate | 0.8 |
| BP270 + pAC7::PyciAB-xylR 0.5 mM glyphosate | 0.726 |
| BP270 + pAC7::PyciAB-xylR 1 mM glyphosate | 0.606 |
The β-xylosidase activity was determined according to script.
| Sample | Start [sec] | End [min] | A420 | A595 | Enzyme activity |
|---|---|---|---|---|---|
| BP270 + pAC7 0 mM glyphosate | 0 | 17 | 0.312 | 0.34 | 107.9 |
| BP270 + pAC7 0.5 mM glyphosate | 0.5 | 17.5 | 0.259 | 0.288 | 105.8 |
| BP270 + pAC7 1 mM glyphosate | 1 | 18 | 0.215 | 0.237 | 106.72 |
| BP270 + pAC7::PyciAB-xylR 0 mM glyphosate | 1.5 | 18.5 | 0.294 | 0.323 | 107 |
| BP270 + pAC7::PyciAB-xylR 0.5 mM glyphosate | 2 | 19 | 0.259 | 0.3 | 101.56 |
| BP270 + pAC7::PyciAB-xylR 1 mM glyphosate | 2.5 | 19.5 | 0.207 | 0.232 | 104.9 |