Display Settings:

Format

Send to:

Choose Destination
Biochem Biophys Res Commun. 2012 Feb 24;418(4):652-6. doi: 10.1016/j.bbrc.2012.01.069. Epub 2012 Jan 24.

High-yield cell-free protein synthesis for site-specific incorporation of unnatural amino acids at two sites.

Author information

  • 1Research School of Chemistry, Australian National University, Canberra, ACT 0200, Australia. ozawa@rsc.anu.edu.au

Abstract

Using aminoacyl-tRNA synthetase/suppressor tRNA pairs derived from Methanocaldococcus jannaschii, an Escherichia coli cell-free protein production system affords proteins with site-specifically incorporated unnatural amino acids (UAAs) in high yields through the use of optimized amber suppressor tRNA(CUA)(opt) and optimization of reagent concentrations. The efficiency of the cell-free system allows the incorporation of trifluoromethyl-phenylalanine using a polyspecific synthetase evolved previously for p-cyano-phenylalanine, and the incorporation of UAAs at two different sites of the same protein without any re-engineering of the E. coli cells used to make the cell-free extract.

Copyright © 2012 Elsevier Inc. All rights reserved.

PMID:
22293204
[PubMed - indexed for MEDLINE]
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Icon for Elsevier Science
    Loading ...
    Write to the Help Desk