Format

Send to

Choose Destination
Methods. 2013 Mar 15;60(1):75-80. doi: 10.1016/j.ymeth.2012.03.008. Epub 2012 Apr 3.

In vitro selection of proteins via emulsion compartments.

Author information

1
Institute of Cellular and Molecule Biology, University of Texas at Austin, TX 78712, USA.

Abstract

In vitro compartmentalization (IVC) is a method to generate numerous, small, aqueous compartments (up to 10(10) compartments per ml) by mixing water, surfactants, and oil. The water phase is surrounded by surfactants and an oil phase, and to a first approximation each water-in-oil compartment is like an artificial cell. By introducing single genes into compartments that are competent for transcription and translation, these cell-like compartments can synthesize RNA protein variants in libraries. Screening or selecting for function has in turn led to schemes for the directed evolution of biomolecules. However, IVC selections can cover larger library sizes, and provide greater control over selection conditions and stringencies. The key issue in designing and executing IVC selections is how to couple genotype and phenotype, and in this review we have organized and presented a variety of mechanisms by which proteins and RNA can attach to or amplify their own templates following emulsification and selection.

PMID:
22491026
DOI:
10.1016/j.ymeth.2012.03.008
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center