Cube E.coli lysates for cell-free protein expression
For increased convenience and higher protein yields (up to 6 mg/ml GFP), the HiYield-T7 lysate was developed. It already includes the T7 RNA polymerase enzyme.
As an alternative to protein expression in living cells, cell-free lysates can be used to obtain recombinant proteins. They are particularly useful for the following applications:
Note that our E.coli lysates require additional components (e.g. amino acids, nucleic acids, DNA polymerase, dialysis devices) that are currently not provided by Cube Biotech. Please refer to the protocols for more details.
In close collaboration with the group of Volker Doetsch and Frank Bernhard, Institute of Biophysics at the University of Frankfurt, Germany, we provide an open system that can be adapted for a range of applications, with dedicated protocols for membrane proteins, continous exchange (dialysis) reactions, and many more. This system takes more work initially than ready to use kits, but provides ultimate flexiblity in reaction variations.
Please note that the E.coli T7 lysates require additional components (e.g. amino acids, nucleic acids, dialysis devices) that are currently not provided by Cube Biotech. Please refer to the protocols for more details.>
Fig. 1: Standard vs. HiYield cell-free expression of GFP. 1,7: MW marker, 2-6: HiYield lysate expression; variation of Mg2+ concentration (14, 16, 18, 20,22 mM). 8-12: Standard lysate expression, same variation of Mg2+ concentrations. A Mg2+ optimum was observed at ca. 16 mM for these combinations of cell-free lysates and other reaction components. Yields range from 1.3-5 mg/ml for HiYield reactions, and from 1-3.8 mg/ml for standard reactions. Data kindly provided by Frank Bernhard, University of Frankfurt, Department of Biophysical Chemistry.