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

被引：29

作者：

Ghadessy, FJ ^{[1
]}

Holliger, P ^{[1
]}

机构：

[1] MRC, Mol Biol Lab, Cambridge CB2 2QH, England

来源：

PROTEIN ENGINEERING DESIGN & SELECTION | 2004年 / 17卷 / 03期

关键词：

compartmentalization; directed evolution; emulsion; rabbit reticulocyte extract; telomerase;

D O I：

10.1093/protein/gzh025

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Emulsion formulations used for in vitro compartmentalization (IVC) methods were found to be 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.

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收藏

页码：201 / 204

页数：4

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