title: Expression of a recombinant elastin-like protein in pichia pastoris.
abstract: 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.
authors.affiliation: Department of Biomedical Engineering, Emory University/Georgia Institute of Technology, Atlanta, GA 30332, USA.
authors.forename: Rory E
authors.initials: RE
authors.lastname: Sallach
authors.forename: Vincent P
authors.initials: VP
authors.lastname: Conticello
authors.forename: Elliot L
authors.initials: EL
authors.lastname: Chaikof
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular / Q000379:methods*; D003062:Codon / Q000235:genetics; D004549:Elastin / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D015723:Gene Library; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000235:genetics; D010843:Pichia / Q000235:genetics* / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D055503:Protein Multimerization; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D020449:Repetitive Sequences, Amino Acid
chemical_list: D003062:Codon; D009842:Oligopeptides; D011994:Recombinant Proteins; D004549:Elastin