Biotechnology Progress · 2009 · 60 citations · 48 references
EngineeringBiological EngineeringBiotechnologyGene ExpressionGenetic EngineeringProtein EngineeringError-prone MutagenesisLibrary ConstructionMutagenesis StrategiesProtein Expression EngineeringProtein Expression OptimizationProtein SynthesisBacterial Protein SynthesisDna Sequence CompositionGene Library SynthesisPremature Translation TerminationRepeat Protein EngineeringSynthesis Of Repetitive Dna Sequences
The translation of highly repetitive gene sequences is often associated with reduced levels of protein expression and may be prone to mutational events. In this report, we describe a modified concatemerization strategy to construct a gene with enhanced sequence diversity that encodes a highly repetitive elastin-like protein polymer for expression in Pichia pastoris. Specifically, degenerate oligonucleotides were used to create a monomer library, which after concatemerization yielded a genetically nonrepetitive DNA sequence that encoded identical pentapeptide repeat sequences. By limiting genetic repetition, the risk of genetic deletions, rearrangements, or premature termination errors during protein synthesis is minimized.
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A monomeric red fluorescent protein
Robert E. Campbell, Oded Tour, Amy E. Palmer et al. · Proceedings of the National Academy of Sciences · 2002 · 2.4K citations · Full text
Planning, Environmental Sciences, Infrastructure Planning +14
Instability of a 550-Base Pair DNA Segment and Abnormal Methylation in Fragile X Syndrome
I. Oberlé, F. Rousseau, Dominique Heitz et al. · Science · 1991 · 1.5K citations
Dan E. Meyer, Ashutosh Chilkoti · Biomacromolecules · 2002 · 551 citations