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Template:UNIPV-Pavia/Project/results/Self-cleaving affinity tags to easily purify proteins
From 2010.igem.org
PHB production
Fusion protein synthesis
These experiments were performed to check bacterial growth and GFP synthesis rate of the following constructs in order to verify right protein folding.
Costitutive promoter devices
Methods
Inoculum (into 5 ml LB+Amp) from glycerol stock of:
- <partinfo>BBa_K300086</partinfo>
- <partinfo>BBa_K300088</partinfo>
- <partinfo>BBa_K300090</partinfo>
- <partinfo>BBa_K300099</partinfo>
- <partinfo>BBa_K173000</partinfo> (positive control)
- <partinfo>BBa_B0031</partinfo> (negative control)
They were let grow ON at +37°C, 220 rpm.
The following day cultures were diluted 1:100 and let grow again for about five hours at +37°C, 220 rpm.
Optical density (O.D.) of each cell culture was than measured with TECAN Infinte F200. Samples were diluted in order to obtain the same O.D. equal to 0,02.
Than we performed a 21 hours' experiment with measurements of absorbance and green fluorescence every five minutes with TECAN Infinite F200. Each value shown is the mean of three measurements, from GFP data that of a non-fluorescent culture (negative control) was subtracted; cultures were shaken for 15 seconds every five minutes.
Results
 |
 |
 |
| Culture | Doubling time [min.] |
|---|---|
| <partinfo>BBa_K173000</partinfo> | 77 |
| <partinfo>BBa_K300086</partinfo> | 74 |
| <partinfo>BBa_K300088</partinfo> | 75 |
| <partinfo>BBa_K300090</partinfo> | 76 |
| <partinfo>BBa_K300099</partinfo> | 76 |
| <partinfo>BBa_B0031</partinfo> | 69 |
HSL inducible devices
 |
 |
 |
| Culture | Doubling time [min.] |
|---|---|
| <partinfo>BBa_K173000</partinfo> | 75 |
| <partinfo>BBa_K300095</partinfo> induced | 121 |
| <partinfo>BBa_K300095</partinfo> not induced | 74 |
| <partinfo>BBa_K300084</partinfo> induced | 123 |
| <partinfo>BBa_K300084</partinfo> not induced | 72 |
| <partinfo>BBa_B0031</partinfo> | 74 |
