Full Description
Despite major advances in HIV treatment, many areas require more study, in order to create efficacious, potent antiretrovirals that can suppress viral load completely and durably without toxic side effects, to define unknown drug targets and fine-tune known targets, and to better understand the interplay between viral and host factors. In "HIV Protocols, Second Edition", expert researchers provide clear, state-of-the-art methods for the study of HIV. Directed toward three specific goals, this text aims to document up-to-date protocols for select aspects of HIV biology, to bring together both virological and immunological approaches in a single, convenient volume, and to present a comprehensive account of a range of techniques not available in any existing HIV protocol book. As a volume in the highly successful Methods in Molecular Biology™ series, the chapters include brief introductions to the subject, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and Notes sections containing priceless tips on troubleshooting and avoiding known pitfalls.
Comprehensive and cutting-edge, "HIV Protocols, Second Edition" is an ideal guide to the wide array of techniques used in fundamental or applied research into the biology and pathogenesis of HIV-1.
Contents
Preparation of Virus Particles and their Analysis.- Methods for Viral RNA Isolation and PCR Amplification for Sequencing of Near Full-Length HIV-1 Genomes.- Purification of HIV-1 Virions by Subtilisin Digestion or CD45 Immunoaffinity Depletion for Biochemical Studies.- Calculating HIV-1 Infectious Titre Using a Virtual TCID50 Method.- Methods To Study Hiv-1 Replication.- Cell-Free Assays for HIV-1 Uncoating.- Real-Time PCR Analysis of HIV-1 Replication Post-entry Events.- Analysis of 2-LTR Circle Junctions of Viral DNA in Infected Cells.- HIV-1 Recombination: An Experimental Assay and a Phylogenetic Approach.- Methods of Preparation and Analysis of Intracellular Reverse Transcription Complexes.- Analysis of Viral and Cellular Proteins in HIV-1 Reverse Transcription Complexes by Co-immunoprecipitation.- Isolation and Analysis of HIV-1 Preintegration Complexes.- Bisarsenical Labeling of HIV-1 for Real-Time Fluorescence Microscopy.- Methods for the Study of HIV-1 Assembly.- Assembly of Immature HIV-1 Capsids Using a Cell-Free System.- Preparation of Recombinant HIV-1 Gag Protein and Assembly of Virus-Like Particles In Vitro.- Methods for the Analysis of HIV-1 Nucleocapsid Protein Interactions with Oligonucleotides.- Methods for Analysis of Incorporation and Annealing Methods for Analysis of tRNALys in HIV-1.- Specialized Approaches To Study Hiv-1 Biology and Pathogenesis.- Somatic Cell Genetic Analyses to Identify HIV-1 Host Restriction Factors.- Rapid, Controlled and Intensive Lentiviral Vector-Based RNAi.- Reverse Two-Hybrid Screening to Analyze Protein-Protein Interaction of HIV-1 Viral and Cellular Proteins.- Methods to Study Monocyte Migration Induced by HIV-Infected Cells.- Novel Mouse Models for Understanding HIV-1 Pathogenesis.- Immunological Studies Of Hiv.-Mucosal Antibody Responses to HIV.- Isolating Mucosal Lymphocytes from Biopsy Tissue for Cellular Immunology Assays.- Quantifying HIV-1-Specific CD8 + T-Cell Responses Using ELISPOT and Cytokine Flow Cytometry.- Multiparameter Flow Cytometry Monitoring of T Cell Responses.- Measuring HIV Neutralization in a Luciferase Reporter Gene Assay.- Assessing the Antiviral Activity of HIV-1-Specific Cytotoxic T Lymphocytes.- Methods for Quantitating Antigen-Specific T Cell Responses Using Functional Assays in Rhesus Macaques.- Drug Resistant Viruses And Viral Evolution.- Isolation of Drug-Resistant Mutant HIV Variants Using Tissue Culture Drug Selection.- Virus Evolution as a Tool to Study HIV-1 Biology.
