Table 1.
eGFP and eGFP variants synthesis as a function of DNA regulatory parts
| Plasmid pBEST- | Plasmid concentration [nM] | Protein production [μM] (mg/ml) | |
|---|---|---|---|
| eGFP | 1 | 0.02 | (0.00067) |
| UTR1-eGFP | 1 | 1.05 | (0.0284) |
| UTR1-eGFP-T500 | 1 | 1.10 | (0.0298) |
| UTR1-eGFP-Del6 | 1 | 3.16 | (0.0836) |
| UTR1-eGFP-Del6-229 | 1 | 3.72 | (0.0944) |
| UTR1-eGFP-Del6-229-T500 | 1 | 4.65 | (0.118) |
| OR2-OR1-Pr-UTR1-eGFP | 1 | 1.40 | (0.0375) |
| OR2-OR1-Pr-UTR1-eGFP-Del6 | 1 | 3.30 | (0.0874) |
| OR2-OR1-Pr-UTR1-eGFP-Del6-229 | 1 | 4.24 | (0.108) |
| OR2-OR1-Pr-UTR1-eGFP-Del6-229-T500 | 1 | 5.07 | (0.129) |
| OR2-OR1-Pr-UTR1-eGFP-Del6-229-T500 | 2 | 10.4 | (0.264) |
| OR2-OR1-Pr-UTR1-eGFP-Del6-229-T500 | 5 | 20.2 | (0.513) |
| OR2-OR1-Pr-UTR1-eGFP-Del6-229-T500 | 10 | 25.2 | (0.639) |
| OR2-OR1-Pr-UTR1-eGFP-Del6-229-T500 | 15 | 24.2 | (0.615) |
| OR2-OR1-Pr-UTR1-eGFP-Del6-229-T500 | 20 | 23.4 | (0.593) |
At 1 nM, protein expression with pBEST-OR2-OR1-Pr-UTR1-eGFP-Del6-229-T500 is about 250 times greater than expression with pBEST-eGFP (conditions: 5 mM magnesium glutamate, 50 mM potassium glutamate, 1.5 mM of each amino acid, 2% PEG 8000). Concentrations of magnesium glutamate and potassium glutamate reported here are the concentrations added to the cell-free reaction. The crude extract, dialyzed against the S30 buffer B, brings an additional 4.5 mM magnesium glutamate and 20 mM potassium glutamate to the cell-free reaction.