UNIT 14B.4 Fluorescently Labeled Human Papillomavirus Pseudovirions for Use in Virus Entry Experiments. Issue 1 (1st May 2015)

• Record Type: Journal Article
• Authors: Samperio Ventayol, Pilar; Schelhaas, Mario
• Is Part Of: Current protocols in microbiology. Volume 37:Issue 1(2015)
• Page Start: 14B.4.1
• Page End: 14B.4.22
• Publication Date: 2015-05-01
• Subjects: HPV -- papillomavirus -- virus entry -- labeling -- microscopy; Microbiology -- Laboratory manuals; Microbiology -- Technique -- Periodicals; Microbiological Techniques -- methods; Containment of Biohazards -- methods; Laboratory Infection -- prevention & control; Microbiology; Microbiology -- Technique; Fulltext; Internet Resources; Laboratory Manuals; Reference Materials; Periodicals; Laboratory manuals; 579.028
• DOI: 10.1002/9780471729259.mc14b04s37
• Languages: English
• ISSNs: 1934-8525

Abstract

Abstract : Human papillomaviruses (HPV) infect skin or mucosal epidermis. The simplistic capsid consists of a major capsid protein L1, a minor capsid protein L2, and a double‐stranded circular DNA of about 8kB in size. The development of HPV‐based vectors [i.e., pseudovirions (PsV)] as tools to study the initial infection has facilitated our understanding of HPV entry. The covalent coupling of fluorescent molecules to these PsV allows following the viruses en route to the nucleus, i.e., the site of replication. In the first section, we describe a facile method to covalently label HPV PsV that retain their infectivity. In this method, fluorophores coupled to a reactive succinimidyl ester are covalently attached to amine residues in the virion in a one‐step chemical reaction. In the second section of this unit, several assays are outlined that use the fluorescently labeled virions for entry studies in live and fixed cells. © 2015 by John Wiley & Sons, Inc.