Fluorescently Labeled Human Papillomavirus Pseudovirions for Use in Virus Entry Experiments
| Autor: | Mario Schelhaas, Pilar Samperio Ventayol |
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| Rok vydání: | 2015 |
| Předmět: |
Infectivity
Staining and Labeling Epidermis (botany) biology viruses Optical Imaging General Medicine Circular DNA Virus Internalization biology.organism_classification Microbiology Virology Molecular biology Optical imaging Capsid Viral entry Humans Parasitology Papillomaviridae Human papillomavirus |
| Zdroj: | Current Protocols in Microbiology |
| ISSN: | 1934-8533 1934-8525 |
| Popis: | 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 8 kB 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. |
| Databáze: | OpenAIRE |
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