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. 2005 Jul 5:4:21.
doi: 10.1186/1475-2859-4-21.

Structural genomics of human proteins--target selection and generation of a public catalogue of expression clones

Konrad Büssow  1 , Christoph Scheich, Volker Sievert, Ulrich Harttig, Jörg Schultz, Bernd Simon, Peer Bork, Hans Lehrach, Udo Heinemann
Affiliations

Structural genomics of human proteins--target selection and generation of a public catalogue of expression clones

Konrad Büssow et al. Microb Cell Fact. .

Abstract

Background: The availability of suitable recombinant protein is still a major bottleneck in protein structure analysis. The Protein Structure Factory, part of the international structural genomics initiative, targets human proteins for structure determination. It has implemented high throughput procedures for all steps from cloning to structure calculation. This article describes the selection of human target proteins for structure analysis, our high throughput cloning strategy, and the expression of human proteins in Escherichia coli host cells.

Results and conclusion: Protein expression and sequence data of 1414 E. coli expression clones representing 537 different proteins are presented. 139 human proteins (18%) could be expressed and purified in soluble form and with the expected size. All E. coli expression clones are publicly available to facilitate further functional characterisation of this set of human proteins.

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Figures

Figure 1
Figure 1
Vector maps. Vector maps of pQStrep2, pQTEV and pSE111
Figure 2
Figure 2
SDS-PAGE of purified human proteins. 15% SDS-PAGE (Coomassie-stained) of proteins expressed in small scale in E. coli and purified by automated immobilised metal chelate affinity chromatography as described in [43]. The identities of the purified proteins are indicated in Table 2. Protein expression was induced at the temperature that is optimal for the individual clone. These temperatures are listed in the supplementary file psfClones.xml. M: Molecular weight marker.
Figure 3
Figure 3
The supplementary XML Additional file 1 file displayed in a web browser.
See this image and copyright information in PMC

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