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Nature BiotechnologyVolume 16, Issue 7, July 1998, Pages 652-656

Man-made cell-like compartments for molecular evolution(Article)

  • aCentre for Protein Engineering, MRC Centre, Hills Rd., Cambridge, CB2 2QH, United Kingdom
  • bLaboratory of Molecular Biology, MRC Centre, Hills Rd., Cambridge, CB2 2QH, United Kingdom

Abstract

Cellular compartmentalization is vital for the evolution of all living organisms. Cells keep together the genes, the RNAs and proteins that they encode, and the products of their activities, thus linking genotype to phenotype. We have reproduced this linkage in the test tube by transcribing and translating single genes in the aqueous compartments of water-in-oil emulsions. These compartments, with volumes close to those of bacteria, can be recruited to select genes encoding catalysts. A protein or RNA with a desired catalytic activity converts a substrate attached to the gene that encodes it to product. In other compartments, substrates attached to genes that do not encode catalysts remain unmodified. Subsequently, genes encoding catalysts are selectively enriched by virtue of their linkage to the product. We demonstrate the linkage of genotype to phenotype in man-made compartments using a model system. A selection for target-specific DNA methylation was based on the resistance of the product (methylated DNA) to restriction digestion. Genes encoding Haelll methyltransferase were selected from a 10- fold excess of genes encoding another enzyme.

Author keywords

Cell-free transcription/translationDirected evolutionDNA methylase

Indexed keywords

EMTREE drug terms:dna methyltransferase
EMTREE medical terms:animal cellarticlecatalysiscell compartmentalizationdna methylationgenetic codegenetic engineeringgenotypemembrane bindingmolecular evolutionnonhumanpriority journal
MeSH:Base SequenceCell CompartmentationDNA MethylationEmulsionsEvolution, MolecularGenotypeModels, GeneticMolecular Sequence DataPhenotypeProtein BiosynthesisProteinsRNASite-Specific DNA Methyltransferase (Cytosine-Specific)Transcription, Genetic
Species Index:Animalia

Chemicals and CAS Registry Numbers:

Emulsions; GGCC-specific DNA methyltransferase, EC 2.1.1.37; Proteins; RNA, 63231-63-0; Site-Specific DNA Methyltransferase (Cytosine-Specific), EC 2.1.1.73

  • ISSN: 10870156
  • CODEN: NABIF
  • Source Type: Journal
  • Original language: English
  • DOI: 10.1038/nbt0798-652
  • PubMed ID: 9661199
  • Document Type: Article

  Tawfik, D.S.; Centre for Protein Engineering, Laboratory of Molecular Biology, MRC Centre, Hills Rd., United Kingdom;
© Copyright 2007 Elsevier B.V., All rights reserved.

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