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This volume details the most updated concepts and experimental protocols developed by leading researchers in the field. Chapters guide readers through methods on bioinformatics tools, hepatitis c virus(HCV) cloning, culture, and purification, HCV life cycle, host immune responses, and small animal models. 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, Hepatitis C Virus Protocols aims to ensure successful results in the further study of this vital field.
Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts
Contenu
Overview of Direct Acting Antiviral Drugs and Drug Resistance of Hepatitis C Virus.- Similarities and Differences between HCV Pseudoparticle (HCVpp) and Cell Culture HCV (HCVcc) in the Study of HCV.- Hepatitis C Virus Database and Bioinformatics Analysis Tools in the Virus Pathogen Resource (ViPR).- Establishment of Replication-competent HCV Strain with Minimum Modifications.- Full-length Open Reading Frame Amplification of Hepatitis C Virus.- InFusion Cloning for the Generation of Biologically Relevant HCV Chimeric Molecular Clones.- Production and Purification of Cell Culture Hepatitis C Virus.- Embryonic or Induced Pluripotent Stem Cell-derived Hepatocellular Systems for HCV Culture.- Molecular Identification of Transmitted/Founder Hepatitis C Viruses and their Progeny by Single Genome Sequencing.- A Cell Culture Model for Persistent HCV Infection.- Expression Cloning of Host Factors Required for the HCV Replication Cycle.- Application of Deep Mutational Scanning in Hepatitis C Virus.- Studying the Hepatitis C Virus-induced Epigenetic Signature after Cure with Direct-acting Antivirals.- Measuring Hepatitis C Virus Envelopment by using a Proteinase K Protection Assay.- Membrane Fusion Assays for Studying Entry Hepatitis C Virus into Cells.- Functional Characterization of Apolipoproteins in the HCV Life Cycle.- Live Cell Imaging of Hepatitis C Virus Trafficking in Hepatocytes.- Cloning and Analysis of Authentic Patient-derived HCV E1/E2 Glycoproteins.- Simultaneous Quantification of Hepatitis C virus Envelope Glycoproteins E1 and E2 by Dual-color Fluorescence Immunoblot Analysis.- Overcoming Challenges of Hepatitis C Virus Envelope Glycoprotein Production in Mammalian Cells.- Monitoring of Interferon Response Triggered by Cells Infected by Hepatitis C Virus or Other Viruses upon Cell-cell Contact.- Complement Regulation and Immune Evasion by Hepatitis C Virus.- Direct ex vivo Functional Analysis of HCV-specific T Cells.- Simultaneous Analysis of HCV-specific CD4 + and CD8 + T Cells by Multi-color Flow Cytometry.- Probing the Antigenicity of HCV Envelope Glycoproteins by Phage Display Antibody Technology. -Isolation of HCV Neutralizing Antibodies by Yeast Display.- Detection of Antibodies to HCV E1E2 by Lectin-Capture ELISA.- In vitro Neutralization Assay using Cultured Hepatitis C Virus.- Standardized Method for the Study of Antibody Neutralization of HCV pseudoparticles (HCVpp).- Detection of Antigen-specific CD8 + T Cells in the Livers of HCV Core Transgenic Mice.- Mice with Chimeric Human Livers and Their Applications.- Mouse Models for Studying HCV Vaccines and Therapeutic Antibodies.