Abstract
Here we present a procedure for preparing amino acid mixtures-having both the desired composition and a physiological pH-at high concentrations for cell-free expression systems. Up to 2.1 mg/mL of active protein was synthesized in batch mode reactions with an all Escherichia coli cell-free expression system. Our method is fast, easy to execute, and economically advantageous compared to expensive commercial kits, making it useful for high-throughput experiments, incorporation of nonstandard amino acids, and cell-free metabolic engineering.
Original language | English (US) |
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Pages (from-to) | 40-43 |
Number of pages | 4 |
Journal | BioTechniques |
Volume | 58 |
Issue number | 1 |
DOIs | |
State | Published - Jan 1 2015 |
Bibliographical note
Publisher Copyright:© 2015, Eaton Publishing Company. All rights reserved.
Keywords
- Amino acid distribution
- Amino acid mixtures
- Cell-free transcription-translation
- pH