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• W1993852129 abstract "In an earlier study, an in vitro evolution procedure was applied to a large population of variants of the Tetrahymena group I ribozyme to obtain individuals with a 10(5)-fold improved ability to cleave a target single-stranded DNA substrate under simulated physiological conditions. The evolved ribozymes also showed a twofold improvement, compared to the wild-type, in their ability to cleave a single-stranded RNA substrate. Here, we report continuation of the in vitro evolution process using a new selection strategy to achieve both enhanced DNA and diminished RNA-cleavage activity. Our strategy combines a positive selection for DNA cleavage with a negative selection against RNA binding. After 36 generations of in vitro evolution, the evolved population showed an approximately 100-fold increase in the ratio of DNA to RNA-cleavage activity. Site-directed mutagenesis experiments confirmed the selective advantage of two covarying mutations within the catalytic core of the ribozyme that are largely responsible for this modified behavior. The population of ribozymes has now undergone a total of 63 successive generations of evolution, resulting in an average of 28 mutations relative to the wild-type that are responsible for the altered phenotype." @default.
• W1993852129 created "2016-06-24" @default.
• W1993852129 creator A5044119161 @default.
• W1993852129 creator A5064029544 @default.
• W1993852129 date "1996-09-01" @default.
• W1993852129 modified "2023-10-16" @default.
• W1993852129 title "Specialization of the DNA-cleaving Activity of a Group I Ribozyme ThroughIn VitroEvolution" @default.
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• W1993852129 doi "https://doi.org/10.1006/jmbi.1996.0496" @default.
• W1993852129 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/8809177" @default.
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