Cloning, expression and functional assessment of elastin like protein: a new protein in medical engineering

Abstract

Introduction: Elastin like protein or ELP is a synthetic biopolymer consisting the pentapeptide repeats of VPGXG (X can be any amino acid except Pro). This protein is the thermal responsive polypeptide that undergoes a reversible phase transition. At a temperature below the transition temperature (Tt), ELP molecules assume an extended conformation and thus are soluble in aqueous solution; but upon the temperature shift higher than the Tt, however, ELPs become insoluble and form the segregated phase prone to ‘aggrigate’ form. The aim of this study was cloning, expression and activity of ELP60 according to Tt. Methods: Firstly, ELP10 (150 bp) was synthesized and cloned into pBluescript. Then, Elp60 (900 bp) was produced using recursive directional ligation (RDL). Finally, ELP60 was subcloned into modified pET25 and ELP60 expression was confirmed using SDS-PAGE method. Also, ELP60 were purified according to Tt and performance of inverse transition cycle (ITC), which confirmed its activity.   Results: The results were shown that the pET-ELP60 constructed and ELP60 expressed; it was successfully purified (about 90%) in one step and non-chromatographic method. Conclusion: From produced recombinant protein can be used for simple and easy purification of the recombinant protein as pharmaceutical drug, smart drug delivery and tissue engineering

Keywords

• Cloning
• Elastin Liked Protein
• Inverse Transition Cycling
• Purification
• Recursive Directional Ligation.