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Directed evolution of an RNA enzyme

Academic Article
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Overview

authors

  • Beaudry, A. A.
  • Joyce, Gerald

publication date

  • July 1992

journal

  • Science  Journal

abstract

  • An in vitro evolution procedure was used to obtain RNA enzymes with a particular catalytic function. A population of 10(13) variants of the Tetrahymena ribozyme, a group I ribozyme that catalyzes sequence-specific cleavage of RNA via a phosphoester transfer mechanism, was generated. This enzyme has a limited ability to cleave DNA under conditions of high temperature or high MgCl2 concentration, or both. A selection constraint was imposed on the population of ribozyme variants such that only those individuals that carried out DNA cleavage under physiologic conditions were amplified to produce "progeny" ribozymes. Mutations were introduced during amplification to maintain heterogeneity in the population. This process was repeated for ten successive generations, resulting in enhanced (100 times) DNA cleavage activity.

subject areas

  • Animals
  • Base Composition
  • Base Sequence
  • Catalysis
  • DNA, Single-Stranded
  • Genotype
  • Hot Temperature
  • Magnesium Chloride
  • Molecular Sequence Data
  • Mutagenesis
  • Mutagenesis, Site-Directed
  • Phenotype
  • Polymerase Chain Reaction
  • RNA, Catalytic
  • Substrate Specificity
  • Tetrahymena thermophila
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Identity

International Standard Serial Number (ISSN)

  • 0036-8075

Digital Object Identifier (DOI)

  • 10.1126/science.1496376

PubMed ID

  • 1496376
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Additional Document Info

start page

  • 635

end page

  • 641

volume

  • 257

issue

  • 5070

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