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This volume provides various techniques and methodologies currently used in the study of MERS-CoV. Chapters are divided into four parts detailing evolution and entry of MERS-coronavirus, genetic alteration and structural determination of MERS-coronavirus proteins, quantitation of virus and anti-viral factors, and mouse models for MERS -coronavirus. Written in the highly 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, MERS Coronavirus: Methods and Protocols aims to ensure successful results in the further study of this vital field.
PART I: Evolution and Entry of MERS-Coronavirus
1. Studying Evolutionary Adaptation of MERS-CoV
Michael Letko and Vincent Munster
2. Evaluating MERS-CoV Entry Pathways
Enya Qing, Michael P Hantak, Gautami G Galpalli, and Tom Gallagher
3. Biochemical Characterization of Middle East Respiratory Syndrome Coronavirus Spike Protein Proteolytic Processing
Gary R. Whittaker and Jean K. Millet
4. Crystallization and Structural Determination of the Receptor-binding Domain of MERS-CoV Spike Glycoprotein
Haixia Zhou, Shuyuan Zhang, and Xinquan Wang
PART II: Genetic Alteration and Structural Determination of MERS-Coronavirus Proteins
5. Bacterial Artificial Chromosome-based Lambda Red Recombination with the I-SceI Homing Endonuclease for Genetic Alteration of MERS-CoV
Anthony R. Fehr
6. Deducing the Crystal Structure of MERS-CoV Helicase
Sheng Cui and Wei Hao
PART III: Quantitation of Virus and Anti-Viral Factors
7. Antigen Capture Enzyme-linked Immunosorbent Assay for Detecting Middle East Respiratory Syndrome Coronavirus in Humans
Joshua Fung, Susanna K. P. Lau, and Patrick C. Y. Woo
8. Quantification of the Middle East Respiratory Syndrome-Coronavirus RNA in Tissues by Quantitative Real-Time RT-PCR
Abdullah Algaissi, Anurodh S. Agrawal, Anwar M. Hashem, and Chien-Te K. Tseng
9. Evaluation of MERS-CoV Neutralizing Antibodies in Sera Using Live Virus Microneutralization Assay
Abdullah Algaissi and Anwar M Hashem
10. Generation of MERS-CoV Pseudotyped Viral Particles for the Evaluation of Neutralizing Antibodies in Mammalian Sera
Abdulrahman Almasaud, Naif Khalaf Alharbi, and Anwar M Hashem
11. Qualitative and Quantitative Determination of MERS-CoV S1-specific Antibodies using ELISA
Sawsan S Al-amri and Anwar M Hashem
PART IV: Mouse Models for MERS –Coronavirus
12. Genetically Engineering a Susceptible Mouse Model for MERS-CoV-Induced
Acute Respiratory Distress Syndrome
Sarah R. Leist and Adam S. Cockrell
13. Development of a Mouse Adapted MERS Coronavirus
Kun Li and Paul B. McCray, Jr.
14. Metabolite, Protein, and Lipid Extraction (MPLEx): A Method that Simultaneously Inactivates Middle East Respiratory Syndrome Coronavirus and Allows Analysis of Multiple Host Cell Components Following Infection
Carrie D. Nicora, Amy C. Sims, Kent J. Bloodsworth, Young-Mo Kim, Ronald J. Moore, Jennifer E. Kyle, Ernesto S. Nakayasu, and Thomas O. Metz
15. Evaluation of Activation and Inflammatory Activity of Myeloid Cells During
Pathogenic Human Coronavirus Infection
Rudragouda Channappanavar and Stanley Perlman
16. Histopathologic Evaluation and Scoring of Viral Lung InfectionDavid K. Meyerholz and Amanda P. Beck
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