Cell-Free Expression Systems

by Endo, Yaeta; Takai, Kazuyuki; Ueda, Takuya

• ISBN13: 9781603273305
• ISBN10: 1603273301
• Format: Hardcover
• Copyright: 2010-01-03
• Publisher: Humana Pr Inc

Summary

During the past decade as the data on gene sequences and expression patterns rapidly accumulated, cell-free protein synthesis technology has also experienced a revolution, becoming a powerful tool for the preparation of proteins for their functional and structural analysis. In Cell-Free Protein Production: Methods and Protocols, experts in the field contribute detailed techniques, the uses of which expand deep into the studies of biochemistry, molecular biology, and biotechnology. Beginning briefly with basic methods and historical aspects, the book continues with thorough coverage of protein preparation methods, the preparation of proteins that are generally difficult to prepare in their functional forms, applications of the cell-free technologies to protein engineering, as well as some methods that are expected to constitute a part of future technologies. Written in the highly successful Methods in Molecular Biology'„¢ series format, the chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.Authoritative and cutting-edge, Cell-Free Protein Production: Methods and Protocols aims to help researchers continue the growth of the vital exploration of cell-free sciences and technologies in order to better understand the dynamic lives of cells.

Table of Contents

Preface	p. v
Contributors	p. ix
Cell-Free Protein Preparation Through Prokaryotic Transcription-Translation Methods	p. 1
PURE Technology	p. 11
The Cell-Free Protein Synthesis System from Wheat Germ	p. 23
A Cell-Free Protein Synthesis System from Insect Cells	p. 31
Cell-Free Protein Synthesis Systems with Extracts from Cultured Human Cells	p. 43
Analysis of Protein Functions Through a Bacterial Cell-Free Protein Expression System	p. 53
Cell-Free-Based Protein Microarray Technology Using Agarose/DNA Microplate	p. 63
An Efficient Approach to the Production of Vaccines Against the Malaria Parasite	p. 73
Protein Engineering Accelerated by Cell-Free Technology	p. 85
Cell-Free Protein Production System with the E. coli Crude Extract for Determination of Protein Folds	p. 101
NMR Assignment Method for Amide Signals with Cell-Free Protein Synthesis System	p. 113
Cell-Free Protein Synthesis Technology in NMR High-Throughput Structure Determination	p. 127
Cell-Free Protein Synthesis for Structure Determination by X-ray Crystallography	p. 149
Production of Multi-Subunit Complexes on Liposome Through an E. coli Cell-Free Expression System	p. 161
Synthesis of a Hetero Subunit RNA Modification Enzyme by the Wheat Germ Cell-Free Translation System	p. 173
Strategies for the Cell-Free Expression of Membrane Proteins	p. 187
Production of Membrane Proteins Through the Wheat-Germ Cell-Free Technology	p. 213
Ribosome Display with the PURE Technology	p. 219
Incorporation of 3-Azidotyrosine into Proteins Through Engineering Yeast Tyrosyl-tRNA Synthetase and Its Application to Site-Selective Protein Modification	p. 227
Synthesis of Functional Proteins Within Liposomes	p. 243
Index	p. 257
Table of Contents provided by Ingram. All Rights Reserved.