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
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.
引用
收藏
页码:201 / 204
页数:4
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