The field of Structural Genomics has produced many technological advances that transform and accelerate structure solution and analysis. Structural Genomics: General Applications emphasizes the benefits to the wider structural research community. It also reflects the current trend in tackling the more ambitious challenges of studying macromolecular machineries and complexes. Divided into three convenient sections, topics include the cloning and production of proteins for structural studies, experimental methods, and computational methods and data analysis. 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, Structural Genomics: General Applications aims primarily to channel spin-off technologies to the average structural biologist in a small or medium-sized laboratory.

InhaltsangabePart I. Cloning, Expression and Protein Production 1. DisMeta - a Meta Server for Construct Design and Optimization             Yuanpeng Janet Huang, Thomas B. Acton, and Gaetano T. Montelione       2. Stable Expression Clones and Auto-Induction for Protein Production in E. Coli            F. William Studier 3. High-Throughput Expression Screening and Purification of Recombinant Proteins in E. coli            Natalie J. Saez and Renaud Vincentelli 4. Medium Throughput Production of Recombinant Human Proteins: Ligation Independent Cloning            Claire Strain-Damerell, Nicola A. Burgess-Brown, Pravin Mahajan, and Opher Gileadi 5. Medium Throughput Production of Recombinant Human Proteins: Protein Production in E. coli            Nicola A. Burgess-Brown, Pravin Mahajan, Claire Strain-Damerell, Opher Gileadi, and Susanne Gräslund 6. Medium Throughput Production of Recombinant Human Proteins: Protein Production in Insect Cells            Pravin Mahajan, Nicola A. Burgess-Brown, Claire Strain-Damerell, and Opher Gileadi 7. OmniBac: Universal Multigene Transfer Plasmids for Baculovirus Expression Vector Systems            Deepak B. Thimiri Govinda Raj, Lakshmi S. Vijayachandran, and Imre Berger 8. Multiprotein Complex Production in Insect Cells by Using Polyproteins            Yan Nie, Itxaso Bellon-Echeverria, Simon Trowitzsch, Christoph Bieniossek, and Imre Berger 9. Expression Screening in Mammalian Suspension Cells             Susan D. Chapple and Michael R. Dyson 10. Cell-free Expression of Protein Complexes for Structural BiologyTakaho Terada, Takeshi Murata, Mikako Shirouzu, and Shigeyuki Yokoyama 11. Cell-Free Protein Synthesis for Functional and Structural Studies            Shin-ichi Makino, Emily T. Beebe, John L. Markley, and Brian G. Fox 12. Insoluble Protein Purification with Sarkosyl: Facts and Precautions            Ben Chisnall, Courtney Johnson, Yavuz Kulaberoglu, and Yu Wai Chen Part II. Experimental Structure Determination and Characterisation 13. Estimation of Crystallization Likelihood Through a Fluorimetric Thermal Stability Assay            Vincent Mariaule, Florine Dupeus, and José A. Márquez 14. CrystalDirect: A Novel Approach for Automated Crystal Harvesting Based on Photoablation of Thin Films             Jose A. Marquez and Florent Cipriani 15. Methods to Refine Macromolecular Structures in Cases of Severe Diffraction Anisotropy             Michael R. Sawaya      16. Applications of NMR-Based PRE and EPR-Based DEER Spectroscopy to Homodimer Chain Exchange Characterization and Structure Determination             Yunhuang Yang, Theresa A. Ramelot, Shuisong Ni, Robert M. McCarrick, and Michael A. Kennedy 17. A Cost-Effective Protocol for the Parallel Production of Libraries of 13CH3-Specifically Labelled Mutants for NMR Studies of High Molecular Weight Proteins            Elodie Crublet, Rime Kerfah, Guillaume Mas, Marjolaine Noirclerc-Savoye, Violaine Lantez, Thierry Vernet, and Jerome Boisbouvier 18. High-Throughput SAXS for the Characterization of Biomolecules in Solution: A Practical Approach            Kevin N. Dyer, Michal Hammel,  Robert P. Rambo, Susan E. Tsutakawa, Ivan Rodic, Scott Classen, John A. Tainer, and Greg L. Hura 19. Measuring Spatial Restraints on Native Protein Complexes using Isotope-Tagged Chemical Cross-Linking and Mass Spectrometry             Franz Herzog Part III. Computational Methods and Structural Data Analyses 20. Modeling of Proteins and Their Assemblies with the Integrative Modeling Platform             Benjamin Webb, Keren Las