Prion Extraction Methods: Comparison of Bead Beating, Ultrasonic Disruption, and Repeated Freeze-Thaw Methodologies for the Recovery of Functional Renilla-Prion Fusion Protein from Bacteria

Abstract

© Springer Science+Business Media New York 2016. Molecular DNA technology allows for production of mammalian proteins in bacteria at sufficient quantities for downstream use and analysis. Variation in design and engineering of DNA expression vectors imparts selective alterations resulting in the generation of fusion proteins with intrinsic reporters or tags used to facilitate their isolation or detection. A multitude of methods are available for the disruption of bacteria that aid in the recovery of recombinant protein. Here we compare three standard methods for the disruption of bacteria (bead beating, repeated freeze-thaw, and ultrasonic disruption) and evaluate the yield of a functional recombinant renilla-prion fusion protein from crude bacterial extracts. This report provides methods and guidance to maximize the yield of recombinant fusion proteins from bacteria for downstream applications.