Document Detail

Expression and purification of ZEBRA fusion proteins and applications for the delivery of macromolecules into mammalian cells.
MedLine Citation:
PMID:  19016434     Owner:  NLM     Status:  MEDLINE    
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 beta-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.
Romy Rothe; Jean-Luc Lenormand
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Current protocols in protein science / editorial board, John E. Coligan ... [et al.]     Volume:  Chapter 18     ISSN:  1934-3663     ISO Abbreviation:  -     Publication Date:  2008 Nov 
Date Detail:
Created Date:  2008-11-21     Completed Date:  2008-12-19     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101287868     Medline TA:  Curr Protoc Protein Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  Unit 18.11     Citation Subset:  IM    
HumProTher Laboratory, TheREx-GREPI, University of Joseph Fourier, UFR de Médecine, La Tronche, France.
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MeSH Terms
Gene Expression
Green Fluorescent Proteins / genetics
Herpesvirus 4, Human / chemistry,  genetics
Recombinant Fusion Proteins / genetics*,  isolation & purification*
Viral Regulatory and Accessory Proteins / genetics
beta-Galactosidase / genetics
Reg. No./Substance:
0/Recombinant Fusion Proteins; 0/Viral Regulatory and Accessory Proteins; 0/enhanced green fluorescent protein; 147336-22-9/Green Fluorescent Proteins; EC

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