Gene cassette design, cloning and expression of recombinant elastin like polypeptide to produce a functional biomaterial in tissue engineering

Abstract

Introduction: Elastin-like polypeptide (ELP) s an artificial biopolymer and a pentapeptide with repeating motif of Val-Pro-Gly-Xaa-Gly (VPGXG). The structure is derived from extracellular cellular matrix (ECM) elastin. Protein-based polymers, which are composed of repeat units of natural or unnatural amino acids, have recently emerged as a promising new class of materials. The unique properties of ELPs such as being genetically encoded in a heterologous host (e.g., bacteria or eukaryotic cell), excitability under the influence of environmental factors, biocompatibility and biodegradability made them popular in a wide variety of biomedical applications. Materials and Methods: In this study, 330bp GRGDS-ELP monomer gene (expressing21 pentapeptide) was oligomerized into the cloning vector pUC to produce 990bp trimer gene (expressing63 fusion pentapeptide).After confirming gene structure by sequencing, the gene sequence was subcloned into pET-MOD expression vector and transduced into the E.coli. Results: Recombinant protein expression was induced by IPTG and the accuracy of the recombinant protein was confirmed by SDS-PAGE and Western blotting. Conclusion: With mentioned method, ELP was obtained for further studies