A time- and cost-efficient system for high-level protein production in mammalian cells

被引：593

作者：

Aricescu, A. Radu ^{[1
]}

Lu, Weixian ^{[1
]}

Jones, E. Yvonne ^{[1
]}

机构：

[1] Univ Oxford, Canc Res UK, Receptor Struct Res Grp, Div Struct Biol,Wellcome Trust Ctr Human Genet, Oxford OX3 7BN, England

来源：

ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY | 2006年 / 62卷

基金：

英国医学研究理事会;

关键词：

D O I：

10.1107/S0907444906029799

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Most proteins for structural biology studies are produced by high- level expression in Escherichia coli. However, prokaryotic based expression systems fail to generate correctly folded functional forms of many proteins and hence a variety of eukaryotic based expression systems have been developed. Of these, yeast and baculovirus- infected insect cells currently represent the expression systems of choice for structural biologists. Here, protocols for a simple, fast and affordable method for transient protein expression in mammalian cells are reported. The results demonstrate that it combines several features necessary for the production of suitable samples for structural biology, in particular protein crystallography, namely high protein yield, straightforward purification, selenomethionine incorporation and control of N- linked glycosylation. The system is suitable for use in conventional laboratories or can be implemented in a medium- or high-throughput pipeline.

引用

页码：1243 / 1250

页数：8

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