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Full Description
Cell-free protein expression promises to narrow the technological gap between DNA and protein technologies and provide a platform for broad application of synthetic biology principles in the Life Sciences. It is a rapid and high throughput methodology for the conversion of DNA encoded genetic information into protein-mediated biochemical activities. Cell-Free Protein Synthesis: Methods and Protocols brings together the key opinion leaders of cell-free technology development and provides case studies and detailed protocols for the application of cell-free methodology. Chapters cover the main directions in the development of cell-free technologies including several recently developed cell-free systems, as well as a number of applications of cell-free systems ranging from discovery of biofuel enzymes to in vitro assembly of viruses. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.
Authoritative and easily accessible, Cell-Free Protein Synthesis: Methods and Protocols seeks to serve a wide variety of scientists with its well-honed methodologies.
Contents
Production of Eukaryotic Cell-Free Lysate from Leishmania tarentolae.- Bioinformatics Analysis and Optimization of Cell-Free Protein Expression.- A Cell-Free Expression Screen to Identify Fusion Tags for Improved Protein Expression.- One-Pot, Microscale Cell-Free Enzyme Expression and Screening.- Cell-Free Translation of Biofuels Enzymes.- Cloning-Independent Expression and Screening of Enzymes using Cell-Free Protein Synthesis Systems.- High Level Cell-Free Production of Membrane Proteins with Nanodiscs.- Cell Free Protein Based Enzyme Discovery and Protein-Ligand Interaction Study.- Human Cell Extract-Derived Cell-Free Systems for Virus Synthesis.- Cell-Free Protein Synthesis in Microfluidic 96-Well Plates.- Preparation of Multiple Site-Specific Mutant Proteins for NMR Studies by PCR-Directed Cell-Free Protein Synthesis.- Site-Specific Incorporation of Unnatural Amino Acids into Proteins by Cell-Free Protein Synthesis.- In vitro Translation of Papillomavirus Authentic and Codon-Modified L1 Capsid Gene mRNAs in Mouse Keratinocyte Cell-Free Lysate.- An Optimized Yeast Cell-Free Lysate System for In vitro Translation of Human Virus mRNA.- In vitro Translation Based Protein Kinase Substrate Identification.- Preparation of Protein Arrays using Cell-Free Protein Expression.- Post-Transcriptional Control of Protein Synthesis in Drosophila S2 Cell-Free System.- Cell-Free Membrane Protein Expression.- The PURE System for Protein Production.- A Cell-Free Protein Synthesis System from Insect Cells.- A Cell-Free Expression Platform for Production of Protein Microarrays.
