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Team:Groningen/Promoters
From 2009.igem.org
[http://2009.igem.org/Team:Groningen http://2009.igem.org/wiki/images/f/f1/Igemhomelogo.png]
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This page shows the measurements we did of our constitutive promoters and derives approximate RPUs relative to the weakest promoter we use. For briefness the lines in the graphs use the original numbering, which can be summarized as follows:
| Plasmid/Part → | [http://partsregistry.org/Part:pSB1AC3 pSB1AC3] | [http://partsregistry.org/Part:pSB3K3 pSB3K3] | [http://partsregistry.org/Part:BBa_J23109 BBa_J23109] | [http://partsregistry.org/Part:BBa_J23106 BBa_J23106] | [http://partsregistry.org/Part:BBa_J23100 BBa_J23100] | control | |||||||
| Medium → | LB | TY | LB | TY | LB | TY | LB | TY | LB | TY | LB | TY | |
| Dilution (of culture) ↓ | 1 | 7 | 2 | 8 | 3 | 9 | 4 | 10 | 5 | 11 | 6 | 12 | |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 100x | A | ||||||||||||
| B | |||||||||||||
| 200x | C | ||||||||||||
| D | |||||||||||||
| 400x | E | ||||||||||||
| F | |||||||||||||
Background
The following graphs show the optical density and fluorescence measurements for the controls. As can be seen 4 of the LB medium controls were probably contaminated, so these are ignored in the rest of the computations.
| LB controls | TY controls |
|---|---|
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Based on the non-contaminated control measurements we find the following average background values:
- LB
- OD600: 0.094262500378219 (A, F)
- Fluorescence: -11.4090909090909 (A)
- Fluorescence: -20.6136363636364 (B,C,D,E,F)
- TY
- OD600: 0.09115113622763 (A-F)
- Fluorescence: -9.06818181818182 (A)
- Fluorescence: -20.05 (B,C,D,E,F)
Using the above background values we get the graphs below.
| BBa_J23109 | BBa_J23106 | BBa_J23100 | |
|---|---|---|---|
| LB | Loading graph...
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| TY | Loading graph...
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| LB | Loading graph...
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| TY | Loading graph...
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Relative promoter units
Using the computed absorbance and fluorescence values we can determine RPUs (Kelly2009) using the formula:
((d/dt)Fθ/ABSθ)/((d/dt)FJ23100/ABSJ23100)
Note that we did not test with the official reference promoter and with RFP instead of GFP, as this happened to be easily avaible. In other words, these results should NOT be compared with other computed RPU values, but only be taken as a rough indication of the relative performance of these three promoters.
| Reference = BBa_J23100 | ||
|---|---|---|
| BBa_J23109 | BBa_J23106 | |
| LB | Loading graph...
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| TY | Loading graph...
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Based on the data from t>400min (>24000s) the minimum, maximum and average RPU values were computed (see the "RPU total" sheet in [http://spreadsheets.google.com/pub?key=tL1vk4bsWObQyq46vPnbV-Q the underlying spreadsheet] for the raw values, before rounding):
| BBa_J23109 | BBa_J23106 | |
|---|---|---|
| Min RPU | -0.011 | 0.28 |
| Max RPU | 0.033 | 0.84 |
| Average RPU | 0.015 | 0.49 |
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