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Table 1 eGFP and eGFP variants synthesis as a function of DNA regulatory parts

From: Efficient cell-free expression with the endogenous E. Coli RNA polymerase and sigma factor 70

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)
  1. 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.