Full Description
Human Retroviruses: Methods and Protocols collects key experimental protocols that have provided the basis of the major discoveries of the field. Split into five sections, this detailed volume covers mapping of the HIV life cycle, isolation, co-receptor use, and cell tropism of HIV-1, in vivo quantification of HIV-1, biological aspects of HIV-1, as well as HTLVs. Some articles explore "assay and function of accessory genes", largely involving the interface between retroviral and host factors, the extracellular role of Tat and Tax, resembling the function of cytokines, and the biotechnological exploitation of HIV as lentiviral vector to carry foreign genes with therapeutic value. 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.
Comprehensive and authoritative, Human Retroviruses: Methods and Protocols provides state-of-art methodological protocols from world leaders in human retrovirology, essential for any lab working this vital field.
Contents
Virion Attachment & Entry: HIV gp120 Env Biotinylation, gp120 Env, or Integrin Ligand Binding Assay.- CryoEM Analysis of Capsid Assembly and Structural Changes Upon Interactions with a Host Restriction Factor, TRIM5α.- The Fate of HIV-1 Capsid: A Biochemical Assay for HIV-1 Uncoating.- The Cyclosporin A Washout Assay to Detect HIV-1 Uncoating in Infected Cells.- Imaging HIV-1 Nuclear Pre-Integration Complexes.- HIV-1 Reverse Transcription.- RNase H: Specificity, Mechanisms of Action, and Antiviral Target.- HIV-1 Chromatin, Transcription, and the Regulatory Protein Tat.- HIV-1 Rev Function and RNA Nuclear-Cytoplasmic Export.- HIV-1 Accessory Proteins: Nef.- HIV-1 Accessory Proteins: VpR.- HIV-1 Accessory Proteins: Vpu and Vif.- SIVSM/HIV-2 Vpx Proteins : Function and Uses in the Infection of Primary Myeloid Cells.- Imaging of HIV Assembly and Release.- HIV-1 Isolation from Infected Peripheral Blood Mononuclear Cells.- Determination of HIV-1 Co-Receptor Usage.- The Macrophage and HIV: Basic Concepts and Methodologies.- HIV Infection of Dendritic Cells.- Histocultures (Tissue Explants) in Human Retrovirology.- Single Copy Quantification of HIV-1 in Clinical Samples.- Quantification of Total HIV1-DNA in Peripheral Blood Mononuclear Cells.- HIV-1 Based Lentiviral Vectors.- Quantification of miRNA by Poly(A)-RT-qPCR Arrays and Verification of Target Sites in HIV-1 Using a One-LTR Infectious Molecular Clone.- Investigating Human T Cell Lymphotropic Retrovirus (HTLV) Tax Function with Molecular and Immunophenotypic Techniques.- Proviral Load Determination of HTLV-1 and HTLV-2 in Patients' Peripheral Blood Mononuclear Cells by Real-Time PCR.- Quantitative Analysis of HumanT-Lymphotropic Virus Type 1 (HTLV-1) Gene Expression Using Nucleo-Cytoplasmic Fractionation and Splice Junction-Specific Real Time RT-PCR (qRT-PCR).
