Efficient incorporation of unnatural amino acids into proteins in Escherichia coli.
basic_science · Level V
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Abstract
We have developed a single-plasmid system for the efficient bacterial expression of mutant proteins containing unnatural amino acids at specific sites designated by amber nonsense codons. In this system, multiple copies of a gene encoding an amber suppressor tRNA derived from a Methanocaldococcus jannaschii tyrosyl-tRNA (MjtRNATyrCUA) are expressed under control of the proK promoter and terminator, and a gene encoding the desired mutant M. jannaschii tyrosyl-tRNA synthetase (MjTyrRS) is expressed under control of a mutant glnS (glnS') promoter.
Medical subject headings
- Amino Acids
- Escherichia coli Proteins
- Gene Expression Regulation, Bacterial
- Genetic Engineering
- Plasmids