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.

Shin and Noireaux Journal of Biological Engineering 2010 4:8   doi:10.1186/1754-1611-4-8