Human Papillomaviruses methods and protocols 1st Edition by Clare Davy, John Doorbar 1588293734 978-1588293732

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ISBN 10: 1588293734

ISBN 13: 978-1588293732

Author: Clare Davy, John Doorbar

Leading basic researchers and clinical scientists describe in detail a wide variety of established and cutting-edge techniques they have developed to study the lifecycle and biological properties of the human papillomavirus. The authors use these readily reproducible methods, ranging from PCR to propagation of HPV in vitro, to detect and type papillomavirus infections, study the papillomavirus lifecycle, and to produce and functionally analyze papillomavirus proteins. The protocols follow the successful Methods in Molecular Medicine™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.

Human Papillomaviruses methods and protocols 1st Table of contents:

Identification of New Papillomavirus Types

Identification of HPV Variants

Histochemical Analysis of Cutaneous HPV-Associated Lesions

Histological Analysis of Cervical Intraepithelial Neoplasia

Detection of Papillomavirus Proteins and DNA in Paraffin-Embedded Tissue Sections

Detection and Quantitation of HPV Gene Expression Using Real-Time PCR

Analysis of p16INK4a and Integrated HPV Genomes as Progression Markers

Use of Biomarkers in the Evaluation of CIN Grade and Progression of Early CIN

HPV DNA Detection and Typing in Cervical Scrapes

HPV DNA Detection and Typing in Inapparent Cutaneous Infections and Premalignant Lesions

Establishing HPV-Containing Keratinocyte Cell Lines From Tissue Biopsies

Using an Immortalized Cell Line to Study the HPV Life Cycle in Organotypic “Raft” Cultures

Differentiation of HPV-Containing Cells Using Organotypic “Raft” Culture or Methylcellulose

Propagation of Infectious, High-Risk HPV in Organotypic “Raft” Culture

Retrovirus-Mediated Gene Transfer to Analyze HPV Gene Regulation and Protein Functions in Organotypic “Raft” Cultures

The HPV Xenograft Severe Combined Immunodeficiency Mouse Model

The Cottontail Rabbit Papillomavirus Model of High-Risk HPV-Induced Disease

Studying the HPV Life Cycle in 3A Trophoblasts and Resulting Pathophysiology

Replication and Encapsidation of Papillomaviruses in Saccharomyces cerevisiae

Analysis of the Regulation of Viral Transcription

Analysis of HPV Transcription by RPA

Analysis of Regulatory Motifs Within HPV Transcripts

Detection of HPV Transcripts by Nested RT-PCR

Analysis of HPV DNA Replication Using Transient Transfection and Cell-Free Assays

Detection and Quantitation of HPV DNA Replication by Southern Blotting and Real-time PCR

Analysis of E7/Rb Associations

Transformation Assays for HPV Oncoproteins

Analysis of Adeno-Associated Virus and HPV Interaction

In Vitro Assays of Substrate Degradation Induced by High-Risk HPV E6 Oncoproteins

Measuring the Induction or Inhibition of Apoptosis by HPV Proteins

Codon Optimization of Papillomavirus Genes

Generation of HPV Pseudovirions Using Transfection and Their Use in Neutralization Assays

Generation and Applications of HPV Pseudovirions Using Vaccinia Virus

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