Expression of a recombinant elastin-like protein in pichia pastoris.
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.
Sallach
Rory E
RE
Department of Biomedical Engineering, Emory University/Georgia Institute of Technology, Atlanta, GA 30332, USA.
Conticello
Vincent P
VP
Chaikof
Elliot L
EL
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
D003062:Codon; D009842:Oligopeptides; D011994:Recombinant Proteins; D004549:Elastin