Proficient Target Selection in Structural Genomics by In Vitro Protein Expression on Gateway Recombination Plasmids

Abstract

The numerous whole genome-sequencing projects of the recent genomic era resulted in identifying a huge number of genes with unknown functions, which are now waiting to be characterized. The bioinformatic annotation of prokaryotic genomes, for instance, identifies most of the genes, but leaves 30%–50% of them without any functional attributes [1]. To face this new challenge, many “functional genomics” and “structural genomics” initiatives have been launched throughout the world. One of the goals of structural genomics is to accelerate the discovery of original protein folds as the basis to better understand the protein folding mechanisms, and, thus, improve the performances of computer programs for ab initio 3-D structure modeling or “threading” of proteins. The 3-D structure determination of proteins of known function usually provides detailed insights into their mechanisms of action at the molecular level. It is now expected that the 3-D structure of proteins of unknown function will, in many cases, reveal their similarity to previously described protein families and will immediately provide functional hints that can then be tested experimentally.