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One plasmid selection system for the rapid evolution of aminoacyl-tRNA synthetases

Academic Article
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Overview

authors

  • Melancon, C. E.
  • Schultz, Peter

publication date

  • July 2009

journal

  • Bioorganic & Medicinal Chemistry Letters  Journal

abstract

  • We have developed a rapid, straightforward, one plasmid dual positive/negative selection system for the evolution of aminoacyl-tRNA synthetases with altered specificities in Escherichia coli. This system utilizes an amber stop codon containing chloramphenicol acetyltransferase/uracil phosphoribosyltransferase fusion gene. We demonstrate the utility of the system by identifying a variant of the Methanococcus jannaschii tyrosyl synthetase from a library of 10(9) variants that selectively incorporates para-iodophenylalanine in response to an amber stop codon.

subject areas

  • Amino Acyl-tRNA Synthetases
  • Chloramphenicol O-Acetyltransferase
  • Codon, Terminator
  • Directed Molecular Evolution
  • Escherichia coli
  • Methanococcus
  • Pentosyltransferases
  • Phenylalanine
  • Plasmids
  • Tyrosine-tRNA Ligase
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Research

keywords

  • Aminoacyl-tRNA synthetase
  • Directed evolution
  • Selection
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Identity

PubMed Central ID

  • PMC2714362

International Standard Serial Number (ISSN)

  • 0960-894X

Digital Object Identifier (DOI)

  • 10.1016/j.bmcl.2009.04.007

PubMed ID

  • 19398201
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Additional Document Info

start page

  • 3845

end page

  • 3847

volume

  • 19

issue

  • 14

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