ArticleProceedings of the National Academy of Sciences of the United States of America1997
Functional and nonfunctional mutations distinguished by random recombination of homologous genes.
Article in Proceedings of the National Academy of Sciences of the United States of America, 1997. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 70 citations in OpenAlex.
- Characterization of tumor evolution by functional clonality and phylogenetics in hepatocellular carcinoma.Communications biology · 2024Article
- Engineering an efficient and bright split Corynactis californica green fluorescent protein.Scientific reports · 2021Article
- Revealing biases inherent in recombination protocols.BMC biotechnology · 2007Article
- Selection of novel vesicular stomatitis virus glycoprotein variants from a peptide insertion library for enhanced purification of retroviral and lentiviral vectors.Journal of virology · 2006Article
- Computational and experimental analysis of DNA shuffling.Proceedings of the National Academy of Sciences of the United States of America · 2003Article
- Applied molecular evolution of O6-benzylguanine-resistant DNA alkyltransferases in human hematopoietic cells.Proceedings of the National Academy of Sciences of the United States of America · 2001Article
- Predicting crossover generation in DNA shuffling.Proceedings of the National Academy of Sciences of the United States of America · 2001Article
- In vitro evolution of thermostable p53 variants.Protein science : a publication of the Protein Society · 1999Article
- Delineation of an evolutionary salvage pathway by compensatory mutations of a defective lysozyme.Protein science : a publication of the Protein Society · 1998Article
- Engineering of a cold-adapted protease by sequential random mutagenesis and a screening system.Applied and environmental microbiology · 1998Article
- Random-priming in vitro recombination: an effective tool for directed evolution.Nucleic acids research · 1998Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We describe a convenient method for distinguishing functional from nonfunctional or deleterious mutations in homologous genes. High fidelity in vitro gene recombination ("DNA shuffling") coupled with sequence analysis of a small sampling of the shuffled library exhibiting the evolved behavior allows identification of those mutations responsible for the behavior in a background of neutral and deleterious mutations. Functional mutations are expected to occur in 100% of the sequenced screened sample; neutral mutations are found in 50% on average, and deleterious mutations do not appear at all. When used to analyze 10 mutations in a laboratory-evolved gene encoding a thermostable subtilisin E, this method rapidly identified the two responsible for the observed protease thermostability; the remaining eight were neutral with respect to thermostability, within the precision of the screening assay. A similar approach, coupled with selection for growth and survival of the host organism, could be used to distinguish adaptive from neutral mutations.
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