RT Journal
A1 Luan, Chi-Hao
A1 Qiu, Shihong
A1 Finley, James B.
A1 Carson, Mike
A1 Gray, Rita J.
A1 Huang, Wenying
A1 Johnson, David
A1 Tsao, Jun
A1 Reboul, Jérôme
A1 Vaglio, Philippe
A1 Hill, David E.
A1 Vidal, Marc
A1 DeLucas, Lawrence J.
A1 Luo, Ming
T1 High-Throughput Expression of C. elegans Proteins
JF Genome Research 
JO Genome Research 
YR 2004 
FD October 15 
VO 14 
IS 10b 
SP 2102 
OP 2110 
DO 10.1101/gr.2520504 
UL http://genome.cshlp.org/content/14/10b/2102.abstract 
AB Proteome-scale studies of protein three-dimensional structures should provide valuable information for both investigating basic biology and developing therapeutics. Critical for these endeavors is the expression of recombinant proteins. We selected Caenorhabditis elegans as our model organism in a structural proteomics initiative because of the high quality of its genome sequence and the availability of its ORFeome, protein-encoding open reading frames (ORFs), in a flexible recombinational cloning format. We developed a robotic pipeline for recombinant protein expression, applying the Gateway cloning/expression technology and utilizing a stepwise automation strategy on an integrated robotic platform. Using the pipeline, we have carried out heterologous protein expression experiments on 10,167 ORFs of C. elegans. With one expression vector and one Escherichia coli strain, protein expression was observed for 4854 ORFs, and 1536 were soluble. Bioinformatics analysis of the data indicates that protein hydrophobicity is a key determining factor for an ORF to yield a soluble expression product. This protein expression effort has investigated the largest number of genes in any organism to date. The pipeline described here is applicable to high-throughput expression of recombinant proteins for other species, both prokaryotic and eukaryotic, provided that ORFeome resources become available.