A systematic approach for testing expression of human full-length proteins in cell-free expression systems.

ABSTRACT:

Background

The growing field of proteomics and systems biology is resulting in an ever increasing demand for purified recombinant proteins for structural and functional studies. Here, we show a systematic approach to successfully express a full-length protein of interest by using cell-free and cell-based expression systems.

Results

In a pre-screen, we evaluated the expression of 960 human full-length open reading frames in Escherichia coli (in vivo and in vitro). After analysing the protein expression rate and solubility, we chose a subset of 87 plasmids yielding no protein product in E. coli in vivo. These targets were subjected to a more detailed analysis comparing a prokaryotic cell-free E. coli system with an eukaryotic wheat germ system. In addition, we determined the e

SUBMITTER: Langlais C 

PROVIDER: S-EPMC2131746 | biostudies-literature | 2007 Oct

REPOSITORIES: biostudies-literature