Wieslaw Kudlicki
Invitrogen Corporation

Federico Katzen
Invitrogen Corporation

Robert Bennett
Invitrogen Corporation

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About this Book

Following its inception in the 1950s, cell-free protein synthesis made a tremendous impact on the basic life sciences. The use of cell-free systems was key to understanding molecular mechanisms underlying one of the most complicated processes found in nature: protein translation. Since this time, aggressive cutting-edge research and stiff commerical competition have driven the development of a variety of systems with increased productivity, improved protein quality and relatively low production costs. As a result, technology has generated myriad applications that have enabled advances in fields as diverse as systems biology, structural biology, and drug discovery. Cell-Free Protein Expression describes and expands upon many of these applications. The volume has been divided into six main sections. In the first section, many of the most popular sources of cell-free lysates are introduced. The second section focuses on extraordinary advances made in the Escherichia coli-based systems that have enabled reconstitution of the entire translational process, incorporation of post-translational modifications, yield increase, and production of functional membrane proteins. This progress extends the usefulness of cell-free systems into structural biology applications described in the third section and high-content platforms like protein microarrays discussed in the fourth section. The final two sections cover the use of cell-free protein expression technologies in the rational design and directed evolution of proteins within the scientific community.

Table of Contents

1. The Role of Cell-Free Rabbit Reticulocyte Expression Systems
in Functional Proteomics
Michele Arduengo, Elaine Schenborn and Robin Hurst

2. Advances in Insect-Based Cell-Free Protein Expression
Uritza von Groll, Stefan Kubick, Helmut Merk, Wolfgang Stiege
and Frank Schäfer

3. Advantages and Applications of the Batch-Formatted E. coli
Cell-Free Expression System
Julia E. Fletcher, Federico Katzen, Shiranthi Keppetipola, Ashley Getbehead
and Wieslaw A. Kudlicki

4. Energetics in Escherichia coli-Based Batch Cell-Free Systems
Kalavathy Sitaraman and Deb K. Chatterjee

5. Disulfide Bond Formation in Bacteria-Based Cell-Free Protein Expression
Aaron R. Goerke and James R. Swartz

6. The PURE System: A Minimal Cell-Free Translation System
Bei-Wen Ying, Yoshihiro Shimizu and Takuya Ueda

7. Cell-Free Expression Approaches for the Production and Characterization
of Membrane Proteins
Daniel Schwarz, Christian Klammt, Alexander Koglin, Florian Durst, Frank Löhr,
Volker Dötsch and Frank Bernhard

8. Cell-Free Expression for Protein NMR
A.J. Shaka

9. Cell-Free Synthesis of Membrane Proteins for X-Ray Crystallography
Julia E. Fletcher, Federico Katzen, Wieslaw Kudlicki, Yen-Ju Chen, Andrew Chen,
Samatha Lieu and Geoffrey Chang

10. Bacterial Cell-Free Expression Systems for High-Throughput
Protein Production
T.V.S. Murthy, Leonardo Brizuela and Joshua LaBaer

11. Cell-Free Protein Synthesis for Protein Microarrays
Gregory A. Michaud, Michael Salcius, Rebecca Martone, Diane Buhr, John C. Duarte,
Jennifer E. McCague, Xiangdong Liu, Michael Samuels, Christine Stalder, James Ball,
Alex Tikhonov, Shiranthi Keppetipola, Wieslaw Kudlicki, James Meegan
and Barry I. Schweitzer

12. Cell-Free Protein Expression Screening and Protein Immobilization Using
Protein Microarrays
Matthew A. Coleman, Paul Hoeprich, Peter Beernink and Julio A. Camarero

13. Cell-Free Protein Expression Labeling with Fluorophores
Jerzy Olejnik

14. Cell-Free Synthesis of Defined Protein Conjugates by Site-Directed
Cotranslational Labeling
Michael Gerrits, Jan Strey, Iris Claußnitzer, Uritza von Groll, Frank Schäfer,
Martina Rimmele and Wolfgang Stiege

15. C-Terminal Labeling of Proteins Using Fluorescently Conjugated
Puromycin Derivatives
Ichiro Tabuchi

16. Translation Engineering and Synthetic Biology
David A. Roth, Liza S.Z. Larsen and G. Wesley Hatfield

17. Accelerated Protein Evolution Using Ribosome Display
Julie Douthwaite, Lutz Jermutus, Ronald Jackson

18. Application of in Vitro Virus (IVV) Technique for High-Throughput
Analysis of Protein-Protein Interactions
Etsuko Miyamoto-Sato and Hiroshi Yanagawa