Structural genomics for Caenorhabditis elegans: high throughput protein expression analysis.
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.
Finley
James B
JB
Center for Biophysical Sciences and Engineering, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
Qiu
Shi-Hong
SH
Luan
Chi-Hao
CH
Luo
Ming
M
D000818:Animals; D001331:Automation / Q000379:methods; D017173:Caenorhabditis elegans / Q000235:genetics* / Q000378:metabolism; D029742:Caenorhabditis elegans Proteins / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification; D002455:Cell Division; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D019295:Computational Biology / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development; D015870:Gene Expression; D023281:Genomics / Q000379:methods; D006639:Histidine / Q000235:genetics / Q000276:immunology; D010957:Plasmids / Q000235:genetics; D020543:Proteome / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods*; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification; D044763:Ubiquitin-Conjugating Enzymes / Q000235:genetics
D029742:Caenorhabditis elegans Proteins; D020543:Proteome; D011994:Recombinant Proteins; C033223:polyhistidine; D006639:Histidine; D044763:Ubiquitin-Conjugating Enzymes; C489359:ubc-1 protein, C elegans