ArticleJournal of structural and functional genomics2006
An automated high-throughput screening method for the identification of high-yield, soluble protein variants using cell-free expression and systematic truncation.
Article in Journal of structural and functional genomics, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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Who cites it
4 citing papers in PubMed.
- BpeB, a major resistance-nodulation-cell division transporter from Burkholderia cenocepacia: construct design, crystallization and preliminary structural analysis.Acta crystallographica. Section F, Structural biology communications · 2018Article
- High-throughput instant quantification of protein expression and purity based on photoactive yellow protein turn off/on label.Protein science : a publication of the Protein Society · 2013Article
- Enhancement of crystallization with nucleotide ligands identified by dye-ligand affinity chromatography.Journal of structural and functional genomics · 2012Article
- Benefits of structural genomics for drug discovery research.Infectious disorders drug targets · 2009Review
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Authors and funding
5 authors.
Funding
Abstract
A highly automated method for rapidly identifying soluble protein variants with good expression yields has been developed. This method is based on a commercially available in vitro protein expression system. It consists of two polymerase chain reactions (PCR) followed by in vitro protein expression and protein quantification by dot blot. The PCR protocols have been improved and optimized to allow automation using commercial fluid handling devices. A PCR primer design program has also been implemented to streamline protein variant design. This automated protocol is highly reliable and has tremendously improved the throughput of expression screening as compared to conventional cell-based methods and manual in vitro methods. We have applied this method to 32 problematic targets from the TB Structural Genomics Consortium. Experimental results of these studies are reported.
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