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Directed evolution of an RNA enzymeAn in vitro evolution procedures was used to obtain RNA enzymes with a particular catalytic function. A population of 10 exp 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.
Document ID
19920068207
Acquisition Source
Legacy CDMS
Document Type
Reprint (Version printed in journal)
Authors
Beaudry, Amber A.
(NASA Headquarters Washington, DC United States)
Joyce, Gerald F.
(Scripps Institution of Oceanography La Jolla, CA, United States)
Date Acquired
August 15, 2013
Publication Date
July 31, 1992
Publication Information
Publication: Science
Volume: 257
Issue: 5070
ISSN: 0036-8075
Subject Category
Life Sciences (General)
Accession Number
92A50831
Funding Number(s)
CONTRACT_GRANT: NAGW-1671
CONTRACT_GRANT: NIH-AI-30882
Distribution Limits
Public
Copyright
Other

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