INCORPORATION OF TWO DIFFERENT NONCANONICAL AMINO ACIDS INTO A SINGLE PROTEIN
申请人:Liu Wenshe
公开号:US20130078671A1
公开(公告)日:2013-03-28
The invention relates to methods, systems, and compositions for the genetic incorporation of a plurality of different noncanonical amino acids into one target protein. The invention provides for multiple, mutually orthogonal aminoacyl-tRNA synthetase/tRNA pairs that suppress two different selector codons engineered into a polynucleotide molecule. By virtue of the suppression of the selector codons, orthogonal aminoacyl-tRNA synthetase/tRNA pairs permit incorporation of their charged noncanonical amino acids into the corresponding positions in the protein. The noncanonical amino acids provide a wide array of functional capabilities. For example, the noncanonical amino acids can provide a reactive pair of moieties that facilitate the study and manipulation of the target protein.
A Facile System for Genetic Incorporation of Two Different Noncanonical Amino Acids into One Protein in<i>Escherichia coli</i>
作者:Wei Wan、Ying Huang、Zhiyong Wang、William K. Russell、Pei-Jing Pai、David H. Russell、Wenshe R. Liu
DOI:10.1002/anie.201000465
日期:2010.4.19
Two's company: Using a wild‐type or evolved PylRS‐pylTUUA pair to suppress ochre mutation and an evolved MjTyrRS‐Mj pair to suppress amber mutation, twodifferent noncanonical aminoacids (NAAs) have been concomitantly incorporated into one protein in E. coli with high efficiency (see picture, with NAAs 1–4; GFP=green‐fluorescent protein).