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Combinatorial library approaches for improving soluble protein expression in Escherichia coli

✍ Scribed by Hart, Darren J. ;Tarendeau, Franck

Book ID
104478173
Publisher
International Union of Crystallography
Year
2005
Tongue
English
Weight
299 KB
Volume
62
Category
Article
ISSN
0907-4449

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✦ Synopsis

High-throughput screening methodologies are already used in structural biology to define efficient protein crystallization and expression conditions. Recently, screening approaches have been extended to the optimization of genetic constructs for improved soluble protein expression. With similarities to the directed evolution strategies used in protein engineering, a target gene encoding a poorly expressed protein is mutated by truncation, fragmentation or point mutation. Rare clones with improved protein expression characteristics are then isolated from the random library using a phenotypic screen or selection. This article reviews the progress in this field and provides a general overview of relevant mutation methods, screens and selections.

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