Key result
A modified concatemerization strategy yielded a genetically nonrepetitive DNA sequence encoding identical pentapeptide repeats, minimizing genetic errors during protein synthesis in Pichia pastoris.
A modified concatemerization strategy using degenerate oligonucleotides successfully limits genetic repetition and minimizes synthesis errors when expressing highly repetitive elastin-like proteins in Pichia pastoris.
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May improve repetitive protein yields in yeast; leaves open validation for biomaterial applications.
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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Sallach et al. (2009) studied this question. Modified concatemerization strategy using degenerate oligonucleotides was evaluated on Creation of a genetically nonrepetitive DNA sequence encoding identical pentapeptide repeat sequences. A modified concatemerization strategy yielded a genetically nonrepetitive DNA sequence encoding identical pentapeptide repeats, minimizing genetic errors during protein synthesis in Pichia pastoris.
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