Abstract
Unnatural amino acid S-(2-nitrobenzyl)cysteine was incorporated into human erythropoietin by using a programmed suppression of nonsense codon in a cell-free protein synthesis system. Several controlling factors affecting the operational efficiency of the suppression were investigated and optimized. The amount of suppressor tRNA and the concentration of Mg2+ were crucial not only for the efficiency but also for the control of the exact suppression. In addition, sorne general optimization factors are reported in order to improve the efficiency in an unnatural amino acid mutagenesis.
Original language | English |
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Pages (from-to) | 344-347 |
Number of pages | 4 |
Journal | Journal of Microbiology and Biotechnology |
Volume | 13 |
Issue number | 3 |
State | Published - Jun 2003 |
Keywords
- Cell-free protein synthesis
- Erythropoietin
- Salt concentration optimization
- Suppressor tRNA
- Unnatural amino acid mutagenesis