Abstract
The structural genomics initiatives have begun with the aim to create a so-called 'basic set library' of protein folds that will be used to improve protein prediction methods. Such a library is thought to require the determination of up to 10,000 new structures, including representative structures of several sequence variants from each protein fold. To meet this goal in a reasonable time frame and cost, automated systems must be utilized to clone and to identify the soluble recombinant proteins contained in multiple genomes. This paper presents such a system, developed using the genome of Caenorhabditis elegans (19,099 genes) as a model eukaryotic organism for structural genomics. This system successfully automates nearly all aspects of recombinant protein expression analysis including subcloning, bacterial growth, recombinant protein expression, protein purification, and scoring protein solubility.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 49-55 |
| Number of pages | 7 |
| Journal | Protein Expression and Purification |
| Volume | 34 |
| Issue number | 1 |
| DOIs | |
| State | Published - Mar 2004 |
Funding
This work was supported by the NIH Grant 1P50-GM62407. We thank R. Gray for performing protein purification and ELISAs and W. Thomas for producing bacteria plates, picking colonies, and running protein gels. We also thank D. Johnson, and Drs. M. Carson and J. Tsao for maintaining our web page, http://sgce.cbse.uab.edu/index.htm . Finally, we thank Drs. M. Vidal, J. Reboul, and P. Vaglio (Dana–Farber Cancer Institute, Boston, MA) for generating the original C. elegans ORFs and helpful discussions about genomics.
Keywords
- Automation
- High throughput screening
- Protein expression
- Structural genomics
ASJC Scopus subject areas
- Biotechnology