Unit

UNIT 18.11 Expression and Purification of ZEBRA Fusion Proteins and Applications for the Delivery of Macromolecules into Mammalian Cells

  1. Romy Rothe,
  2. Jean-Luc Lenormand

Published Online: 1 NOV 2008

DOI: 10.1002/0471140864.ps1811s54

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Rothe, R. and Lenormand, J.-L. 2008. Expression and Purification of ZEBRA Fusion Proteins and Applications for the Delivery of Macromolecules into Mammalian Cells. Current Protocols in Protein Science. 54:18.11:18.11.1–18.11.29.

Author Information

  1. HumProTher Laboratory, TheREx-GREPI, University of Joseph Fourier, UFR de Médecine, La Tronche, France

Publication History

  1. Published Online: 1 NOV 2008
  2. Published Print: NOV 2008

Abstract

The recent development of peptide carriers for efficient and specific delivery of biologically active molecules into mammalian cells represents a major advance in the study of both normal and uncontrolled cell growth. In the past few years, this technology has been successfully applied to the delivery of therapeutic molecules in animal models, and now some of these carriers are available in the clinic for the treatment of some human diseases. This unit describes the production, in a bacterial expression system, of reporter proteins (EGFP and β-galactosidase) fused to a transduction domain of the Epstein-Barr virus ZEBRA protein, as well as purification of the fusion proteins. The purified fusion proteins can be added to any of a large spectrum of mammalian cells and the internalization process measured by flow cytometry and fluorescence microscopy on live cells. Fluorescence microscopy on fixed cells is used to study their intracellular distribution. Curr. Protoc. Protein Sci. 54:18.11.1-18.11.29. © 2008 by John Wiley & Sons, Inc.

Keywords:

  • protein expression;
  • fusion protein;
  • protein delivery;
  • endocytosis;
  • intracellular distribution;
  • therapeutic proteins