Efficient incorporation of unnatural amino acids into proteins in Escherichia coli
Efficient incorporation of unnatural amino acids into proteins in Escherichia coli
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DOI：
10.1038/nmeth864
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发表时间：
2006-04-01
期刊：
影响因子：
48
通讯作者：
Schultz, PG
中科院分区：
文献类型：
--
作者：
Ryu, YH;Schultz, PG
我们已经开发了一种单质粒系统，用于有效的细菌表达在由琥珀无义密码子指定的特定位点含有非天然氨基酸的突变蛋白。在该系统中，在proK启动子和终止子的控制下表达编码源自詹氏甲烷球菌酪氨酰-tRNA的琥珀抑制tRNA（MjtRNA（CUA）（Tyr））的基因的多个拷贝，以及编码所需突变体M的基因。詹氏酪氨酰-tRNA合成酶（MjTyrRS）在突变型glnS（glnS ′）启动子的控制下表达。
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 (MjtRNA(CUA)(Tyr)) 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.