{"title":"Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes","authors":"Samuel S. Porter, Alison A. McBride","doi":"10.1002/cpmc.101","DOIUrl":null,"url":null,"abstract":"<p>This protocol describes the production of human papillomavirus (HPV)–derived quasiviruses. Quasiviruses are infectious particles that are produced in 293TT packaging cells and contain a complete viral genome. We describe methods for infection of primary human keratinocytes with HPV quasiviruses, as well as assays to measure early viral DNA replication and transcription. Published 2020. U.S. Government.</p><p><b>Basic Protocol 1</b>: Transfection, harvest, and isolation of HPV quasiviruses</p><p><b>Alternate Protocol 1</b>: Packaging HPV DNA replicated in 293TT cells</p><p><b>Alternate Protocol 2</b>: Production of higher-purity quasivirus using the “Ripcord” method</p><p><b>Support Protocol 1</b>: Production of HPV minicircles</p><p><b>Support Protocol 2</b>: Production of recircularized HPV genomes</p><p><b>Support Protocol 3</b>: Screening of fractions for viral proteins</p><p><b>Support Protocol 4</b>: Screening of fractions for viral DNA</p><p><b>Support Protocol 5</b>: Measuring viral titer</p><p><b>Support Protocol 6</b>: Quantitation of quasivirions</p><p><b>Basic Protocol 2</b>: Infection of primary human foreskin keratinocytes with quasivirus</p><p><b>Basic Protocol 3</b>: HPV quasivirus transcription assay</p><p><b>Basic Protocol 4</b>: HPV quasivirus replication assay</p>","PeriodicalId":39967,"journal":{"name":"Current Protocols in Microbiology","volume":"57 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2020-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmc.101","citationCount":"7","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Microbiology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpmc.101","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 7
Abstract
This protocol describes the production of human papillomavirus (HPV)–derived quasiviruses. Quasiviruses are infectious particles that are produced in 293TT packaging cells and contain a complete viral genome. We describe methods for infection of primary human keratinocytes with HPV quasiviruses, as well as assays to measure early viral DNA replication and transcription. Published 2020. U.S. Government.
Basic Protocol 1: Transfection, harvest, and isolation of HPV quasiviruses
Alternate Protocol 1: Packaging HPV DNA replicated in 293TT cells
Alternate Protocol 2: Production of higher-purity quasivirus using the “Ripcord” method
Support Protocol 1: Production of HPV minicircles
Support Protocol 2: Production of recircularized HPV genomes
Support Protocol 3: Screening of fractions for viral proteins
Support Protocol 4: Screening of fractions for viral DNA
Support Protocol 5: Measuring viral titer
Support Protocol 6: Quantitation of quasivirions
Basic Protocol 2: Infection of primary human foreskin keratinocytes with quasivirus
期刊介绍：
Current Protocols in Microbiology provides detailed, step-by-step instructions for analyzing bacteria, animal and plant viruses, fungi, protozoans and other microbes. It offers updated coverage of emerging technologies and concepts, such as biofilms, quorum sensing and quantitative PCR, as well as proteomic and genomic methods. It is the first comprehensive source of high-quality microbiology protocols that reflects and incorporates the new mandates and capabilities of this robust and rapidly evolving discipline.
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