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PEDS Advance Access published online on February 27, 2004
Protein Engineering Design and Selection, doi:10.1093/protein/gzh025
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Received January 22, 2004
Accepted February 18, 2004
Oxford University Press

Communication

A novel emulsion mixture for in vitro compartmentalisation of transcription and translation in the rabbit reticulocyte system

Farid J. Ghadessy 1 and Philipp Holliger 1*

1 MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK


  Abstract

Emulsion formulations used for in vitro compartmentalisation (IVC) methods were found incompatible with protein expression in the rabbit reticulocyte (RRL) system, causing rapid discoloration and translation shutdown. Here we identify possible causes and describe a novel water-in-oil emulsion which abolished discoloration and allowed high-level in-emulsion expression of active luciferase and human telomerase using the RRL. This novel emulsion greatly expands the range of potential protein targets for IVC.

Keywords: directed evolution / compartmentalisation / rabbit reticulocyte extract / emulsion / telomerase


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