Preparing Genes for Repetitive Elastin-Like Polypeptides Using Gibson Assembly.
Elastin-like polypeptides (ELPs) are stimuli-responsive polymers consisting of peptide repeat units from the elastin protein. Most commonly these materials are biosynthesized in E. coli using genes that have been built by standard molecular biology techniques. The inherent repetitive nature of these polypeptides makes the construction of genes that encode them more challenging than for typical recombinant proteins. Described here is a robust technique to produce genes encoding long chains of ELP through successive rounds of Gibson assembly that nearly double the ELP length with each cloning iteration. Using Gibson assembly to produce the genes for high molecular weight ELPs requires fewer steps and significantly less time than current techniques. This approach is modular and allows for the flexible assembly of ELPs with various compositions. The procedure can be adapted for preparing genes encoding other repetitive proteins.
['Deyling|James K|JK|', 'Turk|Edward M|EM|', 'Holland|Nolan B|NB|']
D000595:Amino Acid Sequence D001483:Base Sequence D004549:Elastin D015870:Gene Expression D005822:Genetic Vectors D008970:Molecular Weight D010455:Peptides D015202:Protein Engineering D011994:Recombinant Proteins D012091:Repetitive Sequences, Nucleic Acid
Elastin-like polypeptides Gibson assembly Molecular biology High molecular weight proteins Ligation Repetitive sequence proteins