A high throughput platform for eukaryotic genes.
The objective of structural proteomics is to determine the structures of all protein folds found in nature and develop a public resource to organize and analyze protein structures and fold families. High throughput (HTP) methods, which can process multiple samples in parallel, saving both time and cost, play important roles in achieving this goal. Using C. elegans and human as model organisms, a HTP cloning and expression pipeline was developed for structural proteomics that required production of a large number of recombinant proteins, applying the Gateway cloning/expression technology and utilizing a stepwise automation strategy on an integrated robotic platform. This system can process up to 384 unique samples in parallel and successfully automates most aspects of gene cloning and protein expression analysis, from PCR to protein solubility profiling.
Chen
Yunjia
Y
Department of Microbiology, University of Alabama at Birmingham, Birmingham, AL, USA.
Qiu
Shihong
S
Luan
Chi-Hao
CH
Luo
Ming
M
D000818:Animals; D017173:Caenorhabditis elegans / Q000502:physiology*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D029742:Caenorhabditis elegans Proteins; D011994:Recombinant Proteins