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Team:Warsaw/ExpressionAdaptors/Results
From 2011.igem.org
Results
Using our software tools we have evolved a pool of candidate expression adaptors from which we have picked six sequences (BBa_K528015 to BBa_K528020)with variable expression levels and synthesized them. This way we have obtained expression adaptors on pSB1C3. All adaptors are under BBa_J23102 promoter in order to measure their strength acording to Synthetic Biology standards.
The adaptors have following predicted strengths according to RBSDesigner software:
| Part Name | Description | Expression level in thousands of absolute RBSDesigner units | Expression level as %B0034+GFP |
|---|---|---|---|
| BBa_K528015 | Expression Adapters s00_45k | 45 | 64,29% |
| BBa_K528016 | Expression Adapters s01_24k | 24 | 34,29% |
| BBa_K528017 | expression adapters s02_20k | 20 | 28,57% |
| BBa_K528018 | Expression Adapters s06_16k | 16 | 22,86% |
| BBa_K528019 | Expression Adapters s05_12k | 12 | 17,14% |
| BBa_K528020 | expression adapters s03_65k | 65 | 92,86% |
| BBa_K528021 | BBa_K528018+YFP measurement device | 16 | 22,86% |
| reference | BBa_J23102+B0034+GFP (100% reference construct) | 70 | 100,00% |
Wet lab results
So far we have managed to obtain only BBa_K528021 (BBa_K528018+YFP measurement device) as working part. We have characterized it using fluorimetry in order to see how accurate are RBSCalculator predictions for the BBa_K528018 adaptor. We have obtained following results:
| Part Name | Description | Expression level as %B0034+GFP | Std Dev. |
|---|---|---|---|
| BBa_K528021 | BBa_K528018+YFP measurement device | 20,37% | 3,21% |
| reference | Bba_J23102+B0034+GFP (100% reference construct) | 100,00% | 2,61% |
As you can see prediction is quite accurate. Clonning of other fluorescent proteins and testing the rest of expression adaptors is still to be done.
