DNA fragmentation-based combinatorial approaches to soluble protein expression: Part I. Generating DNA fragment libraries

Prodromou, Chrisostomos, Savva, Renos and Driscoll, Paul C (2007) DNA fragmentation-based combinatorial approaches to soluble protein expression: Part I. Generating DNA fragment libraries. Drug Discovery Today, 12 (21-22). pp. 931-938. ISSN 1359-6446

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Abstract

In addressing a new drug discovery target, the generation of tractable protein substrates for functional and structural analyses can represent a significant hurdle. Traditional approaches rely on protein expression trials of multiple variants in various systems, frequently with limited success. The increasing knowledge base derived from genomics and structural proteomics initiatives assists the bioinformatics-led design of these experiments. Nevertheless, for many eukaryotic polypeptides, particularly those with relatively few homologues, the generation of useful protein products can still be a major challenge. This review describes the basis of efforts to forge an alternative 'domain-hunting' paradigm, based upon combinatorial sampling of expression construct libraries derived by fragmentation of the encoding DNA template, namely the methods and considerations in generating fragment length DNA from target genes. An accompanying review focuses upon the expression screening of such combinatorial DNA libraries for the sampling of the corresponding set of protein fragments.

Item Type: Article
Keywords: Combinatorial Chemistry Techniques/*methods *DNA Fragmentation Drug Design *Gene Library Polymerase Chain Reaction Proteins/*chemistry
Schools and Departments: School of Life Sciences > Biochemistry
Subjects: Q Science > QD Chemistry > QD0241 Organic chemistry > QD0415 Biochemistry
T Technology > TP Chemical technology > TP0248.13 Biotechnology
Depositing User: Chrisostomos Prodromou
Date Deposited: 24 Feb 2015 17:49
Last Modified: 25 Feb 2015 09:28
URI: http://sro.sussex.ac.uk/id/eprint/44366
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