Link A James, Tirrell David A
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125, USA.
Methods. 2005 Jul;36(3):291-8. doi: 10.1016/j.ymeth.2005.04.005.
The genetic code maps one or more of the 61 sense codons onto a set of 20 canonical amino acids. Reassignment of sense codons to non-canonical amino acids in model organisms such as Escherichia coli has been achieved through manipulation of the cellular protein synthesis machinery. Specifically, control of amino acid pools, coupled with engineering of the aminoacyl-tRNA synthetase activity of the host, has enabled assignment of sense codons to a wide variety of non-canonical amino acids under conditions routinely used for expression of recombinant proteins. Codon reassignment is leading to important advances in protein engineering and bioorganic chemistry. Here we summarize some of those advances, and provide detailed protocols for codon reassignment.
遗传密码将61种有义密码子中的一种或多种对应到一组20种标准氨基酸上。通过操纵细胞蛋白质合成机制，已在诸如大肠杆菌等模式生物中实现了将有义密码子重新分配给非标准氨基酸。具体而言，对氨基酸库的控制，再加上对宿主氨酰-tRNA合成酶活性的改造，使得在常规用于重组蛋白表达的条件下，能够将有义密码子分配给多种非标准氨基酸。密码子重新分配正推动蛋白质工程和生物有机化学取得重要进展。在此，我们总结其中的一些进展，并提供密码子重新分配的详细方案。
