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Hepatitis B Virus
Methods and Protocols (Methods in Molecular Biology)
By Haitao Guo (Edited by), Andrea Cuconati (Edited by)

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Format
Hardback, 297 pages
Other Formats Available

Paperback : $373.00

Published
United States, 15 December 2016

This volume serves as a reference for the dissemination of advances made in the study of Hepatitis B Virus (HBV). Hepatitis B Virus: Methods and Protocols details protocols and techniques ranging from cell culture studies to in vivo and clinical immunology. The chapters in this book discuss treatments of in vitro infection systems, analysis and quantification of cccDNA and its mutations; in vitro polymerase activity assays; cellular trafficking of core proteins; intracellular calcium metabolism; detection, cloning, and sequencing of HBV markers; and new strategies aimed at exploiting new mechanisms for drug discovery. The book also covers classical methods for resolution of extracellular viral particles by native gel electrophoresis, and methods for detecting HBV antigens in drug discovery. 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.



Cutting-edge and comprehensive, Hepatitis B Virus: Methods and Protocols is a valuable tool for researchers to use toward their advanced studies in HBV.









Preface... Table of Contents... Contributing Authors...


1. NTCP-Reconstituted In Vitro HBV Infection System Yinyan Sun, Yonghe Qi, Bo Peng, and Wenhui Li


2. Hepatitis B Virus Infection of HepaRG Cells, HepaRG-hNTCP Cells and Primary Human Hepatocytes Yi Ni and Stephan Urban


3. Live Cell Imaging Confocal Microscopy Analysis of HBV Myr-PreS1 Peptide Binding and Uptake in NTCP-GFP Expressing HepG2 Cells Alexander König and Dieter Glebe


4. Intracytoplasmic Transport of Hepatitis B Virus Capsids Quentin Osseman and Michael Kann

5. A Homokaryon Assay for Nucleocytoplasmic Shuttling Activity of HBV Core Protein Ching-Chun Yang, Hung-Cheng Li, and Chiaho Shih


6. Analyses of HBV cccDNA Quantification and Modification Yuchen Xia, Daniela Stadler, Chunkyu Ko, and Ulrike Protzer


7. Detection of HBV cccDNA Methylation from Clinical Samples by Bisulfite Sequencing and Methylation Specific PCR Yongmei Zhang, Richeng Mao, Haitao Guo, and Jiming Zhang


8. A T7 Endonuclease I Assay to Detect TALEN-Mediated Mutation of HBV cccDNA Kristie Bloom, Abdullah Ely, and Patrick Arbuthnot

9. Detection of Hepatocyte Clones Containing Integrated Hepatitis B Virus DNA using Inverse Nested PCR Thomas Tu and Allison R. Jilbert


10. Highly Sensitive Detection of HBV RNA in Liver Tissue by In Situ Hybridization Diego Calabrese and Stefan F. Wieland


11. Immunofluorescent Staining for the Detection of the Hepatitis B Core Antigen in Frozen Liver Sections of Human Liver Chimeric Mice Lena Allweiss, Marc Lütgehetmann, and Maura Dandri


12. Measuring Changes in Cytosolic Calcium Levels in HBV- and HBx-Expressing Cultured Primary Hepatocytes Jessica C. Casciano and Michael J. Bouchard


13. In Vitro Assays for RNA Binding and Protein Priming of Hepatitis B Virus Polymerase Daniel N. Clark, Scott A. Jones, and Jianming Hu


14. In Vitro Enzymatic and Cell Culture-Based Assays for Measuring Activity of HBV RNaseH Inhibitors Elena Lomonosova and John E. Tavis


15. Detection of Hepatitis B Virus Particles Released from Cultured Cells by Particle Gel Assay Ran Yan, Dawei Cai, Yuanjie Liu, and Haitao Guo


16. Microtiter-Format Assays for HBV Antigen Quantitation in Non-Clinical Applications Cally Goddard, Lale-Bildrici Ertekin, Xiahoe Wang, and Andrea Cuconati

17. Deep Sequencing of the Hepatitis B Virus Genome: Analysis of Multiple Samples by Implementation of the Illumina Platform Quan-Xin Long, Jie-Li Hu, and Ai-Long Huang


18. Generation of Replication Competent Hepatitis B Virus Genome from Blood Samples for Functional Characterization Yanli Qin, Yong-Xiang Wang, Jiming Zhang, Jisu Li, and Shuping Tong


19. Hydrodynamic HBV Transfection Mouse Model Li-Ling Wu, Hurng-Yi Wang, and Pei-Jer Chen


20. An ELISPOT-Based Assay to Measure HBV-Specific CD8+ T Cell Responses in Immunocompetent Mice Tracy D. Reynolds, Safiehkhatoon Moshkani, and Michael D. Robek


21. Advanced Method for Isolation of Mouse Hepatocytes, Liver Sinusoidal Endothelial Cells and Kupffer Cells Jia Liu, Xuan Huang, Melanie Werner, Ruth Broering, Dongliang Yang, and Mengji Lu


22. Partial Hepatectomy and Castration of HBV Transgenic Mice Yongjun Tian and Jing-hsiung James Ou


23. Studying HBV Infection and Therapy in Immune Deficient NOD-Rag1-/-IL2RgammaC-null (NRG) Fumarylacetoacetate Hydrolase (Fah) Knockout Mice Transplanted with Human Hepatocytes Feng Li, Kouki Nio, Fumihiko Yasui, Christopher M. Murphy, and Lishan Su


24. Measurement of Antiviral Effect and Innate Immune Response during Treatment of Primary Woodchuck Hepatocytes Marta G. Murreddu, Manasa Suresh, Severin O. Gudima, and Stephan Menne

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Product Description

This volume serves as a reference for the dissemination of advances made in the study of Hepatitis B Virus (HBV). Hepatitis B Virus: Methods and Protocols details protocols and techniques ranging from cell culture studies to in vivo and clinical immunology. The chapters in this book discuss treatments of in vitro infection systems, analysis and quantification of cccDNA and its mutations; in vitro polymerase activity assays; cellular trafficking of core proteins; intracellular calcium metabolism; detection, cloning, and sequencing of HBV markers; and new strategies aimed at exploiting new mechanisms for drug discovery. The book also covers classical methods for resolution of extracellular viral particles by native gel electrophoresis, and methods for detecting HBV antigens in drug discovery. 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.



Cutting-edge and comprehensive, Hepatitis B Virus: Methods and Protocols is a valuable tool for researchers to use toward their advanced studies in HBV.









Preface... Table of Contents... Contributing Authors...


1. NTCP-Reconstituted In Vitro HBV Infection System Yinyan Sun, Yonghe Qi, Bo Peng, and Wenhui Li


2. Hepatitis B Virus Infection of HepaRG Cells, HepaRG-hNTCP Cells and Primary Human Hepatocytes Yi Ni and Stephan Urban


3. Live Cell Imaging Confocal Microscopy Analysis of HBV Myr-PreS1 Peptide Binding and Uptake in NTCP-GFP Expressing HepG2 Cells Alexander König and Dieter Glebe


4. Intracytoplasmic Transport of Hepatitis B Virus Capsids Quentin Osseman and Michael Kann

5. A Homokaryon Assay for Nucleocytoplasmic Shuttling Activity of HBV Core Protein Ching-Chun Yang, Hung-Cheng Li, and Chiaho Shih


6. Analyses of HBV cccDNA Quantification and Modification Yuchen Xia, Daniela Stadler, Chunkyu Ko, and Ulrike Protzer


7. Detection of HBV cccDNA Methylation from Clinical Samples by Bisulfite Sequencing and Methylation Specific PCR Yongmei Zhang, Richeng Mao, Haitao Guo, and Jiming Zhang


8. A T7 Endonuclease I Assay to Detect TALEN-Mediated Mutation of HBV cccDNA Kristie Bloom, Abdullah Ely, and Patrick Arbuthnot

9. Detection of Hepatocyte Clones Containing Integrated Hepatitis B Virus DNA using Inverse Nested PCR Thomas Tu and Allison R. Jilbert


10. Highly Sensitive Detection of HBV RNA in Liver Tissue by In Situ Hybridization Diego Calabrese and Stefan F. Wieland


11. Immunofluorescent Staining for the Detection of the Hepatitis B Core Antigen in Frozen Liver Sections of Human Liver Chimeric Mice Lena Allweiss, Marc Lütgehetmann, and Maura Dandri


12. Measuring Changes in Cytosolic Calcium Levels in HBV- and HBx-Expressing Cultured Primary Hepatocytes Jessica C. Casciano and Michael J. Bouchard


13. In Vitro Assays for RNA Binding and Protein Priming of Hepatitis B Virus Polymerase Daniel N. Clark, Scott A. Jones, and Jianming Hu


14. In Vitro Enzymatic and Cell Culture-Based Assays for Measuring Activity of HBV RNaseH Inhibitors Elena Lomonosova and John E. Tavis


15. Detection of Hepatitis B Virus Particles Released from Cultured Cells by Particle Gel Assay Ran Yan, Dawei Cai, Yuanjie Liu, and Haitao Guo


16. Microtiter-Format Assays for HBV Antigen Quantitation in Non-Clinical Applications Cally Goddard, Lale-Bildrici Ertekin, Xiahoe Wang, and Andrea Cuconati

17. Deep Sequencing of the Hepatitis B Virus Genome: Analysis of Multiple Samples by Implementation of the Illumina Platform Quan-Xin Long, Jie-Li Hu, and Ai-Long Huang


18. Generation of Replication Competent Hepatitis B Virus Genome from Blood Samples for Functional Characterization Yanli Qin, Yong-Xiang Wang, Jiming Zhang, Jisu Li, and Shuping Tong


19. Hydrodynamic HBV Transfection Mouse Model Li-Ling Wu, Hurng-Yi Wang, and Pei-Jer Chen


20. An ELISPOT-Based Assay to Measure HBV-Specific CD8+ T Cell Responses in Immunocompetent Mice Tracy D. Reynolds, Safiehkhatoon Moshkani, and Michael D. Robek


21. Advanced Method for Isolation of Mouse Hepatocytes, Liver Sinusoidal Endothelial Cells and Kupffer Cells Jia Liu, Xuan Huang, Melanie Werner, Ruth Broering, Dongliang Yang, and Mengji Lu


22. Partial Hepatectomy and Castration of HBV Transgenic Mice Yongjun Tian and Jing-hsiung James Ou


23. Studying HBV Infection and Therapy in Immune Deficient NOD-Rag1-/-IL2RgammaC-null (NRG) Fumarylacetoacetate Hydrolase (Fah) Knockout Mice Transplanted with Human Hepatocytes Feng Li, Kouki Nio, Fumihiko Yasui, Christopher M. Murphy, and Lishan Su


24. Measurement of Antiviral Effect and Innate Immune Response during Treatment of Primary Woodchuck Hepatocytes Marta G. Murreddu, Manasa Suresh, Severin O. Gudima, and Stephan Menne

Show more
Product Details
EAN
9781493966981
ISBN
1493966987
Other Information
28 Tables, color; 31 Illustrations, color; 26 Illustrations, black and white; XII, 297 p. 57 illus., 31 illus. in color.
Dimensions
25.4 x 17.8 x 1.9 centimetres (7.32 kg)

Table of Contents

NTCP-Reconstituted In Vitro HBV Infection System.- Hepatitis B Virus Infection of HepaRG Cells, HepaRG-hNTCP Cells and Primary Human Hepatocytes.- Live Cell Imaging Confocal Microscopy Analysis of HBV Myr-PreS1 Peptide Binding and Uptake in NTCP-GFP Expressing HepG2 Cells.- Intracytoplasmic Transport of Hepatitis B Virus Capsids.- A Homokaryon Assay for Nucleocytoplasmic Shuttling Activity of HBV Core Protein.- Analyses of HBV cccDNA Quantification and Modification.- Detection of HBV cccDNA Methylation from Clinical Samples by Bisulfite Sequencing and Methylation Specific PCR.- A T7 Endonuclease I Assay to Detect TALEN-Mediated Mutation of HBV cccDNA.- Detection of Hepatocyte Clones Containing Integrated Hepatitis B Virus DNA using Inverse Nested PCR.- Highly Sensitive Detection of HBV RNA in Liver Tissue by In Situ Hybridization.- Immunofluorescent Staining for the Detection of the Hepatitis B Core Antigen in Frozen Liver Sections of Human Liver Chimeric Mice.- Measuring Changes in Cytosolic Calcium Levels in HBV- and HBx-Expressing Cultured Primary Hepatocytes.- In Vitro Assays for RNA Binding and Protein Priming of Hepatitis B Virus Polymerase.- In Vitro Enzymatic and Cell Culture-Based Assays for Measuring Activity of HBV RNaseH Inhibitors.- Detection of Hepatitis B Virus Particles Released from Cultured Cells by Particle Gel Assay.- Microtiter-Format Assays for HBV Antigen Quantitation in Non-Clinical Applications.- Deep Sequencing of the Hepatitis B Virus Genome: Analysis of Multiple Samples by Implementation of the Illumina Platform.- Generation of Replication Competent Hepatitis B Virus Genome from Blood Samples for Functional Characterization.- Hydrodynamic HBV Transfection Mouse Model.- An ELISPOT-Based Assay to Measure HBV-Specific CD8+ T Cell Responses in Immunocompetent Mice.- Advanced Method for Isolation of Mouse Hepatocytes, Liver Sinusoidal Endothelial Cells andKupffer Cells.- Partial Hepatectomy and Castration of HBV Transgenic Mice.- Studying HBV Infection and Therapy in Immune Deficient NOD-Rag1-/-IL2RgammaC-null (NRG) Fumarylacetoacetate Hydrolase (Fah) Knockout Mice Transplanted with Human Hepatocytes.- Measurement of Antiviral Effect and Innate Immune Response during Treatment of Primary Woodchuck Hepatocytes.

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