Preparing Genes for Repetitive Elastin-Like Polypeptides Using Gibson Assembly.

Abstract

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.