Analyzing the Human Papillomavirus (HPV) Life Cycle in Primary Keratinocytes with a Quantitative Colony-Forming Assay

Michael J Lace; Lubomir P Turek; James R Anson; Thomas H Haugen

doi:10.1002/9780471729259.mc14b02s33

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Journal article

Analyzing the Human Papillomavirus (HPV) Life Cycle in Primary Keratinocytes with a Quantitative Colony-Forming Assay

Michael J Lace, Lubomir P Turek, James R Anson and Thomas H Haugen

Current protocols in microbiology (Online), Vol.33(1), pp.14B.2.1-14B.2.13

05/01/2014

DOI: 10.1002/9780471729259.mc14b02s33

PMID: 24789595

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Abstract

Papillomavirus genomes replicate as extrachromosomal plasmids within infected keratinocytes, requiring the regulated expression of early viral gene products to initially amplify the viral genomes and subvert cell growth checkpoints as part of a complex path to immortalization. Building on contemporary keratinocyte transfection and culture systems, the methods described in this unit form a detailed approach to analyzing critical events in the human papillomavirus (HPV) life cycle, utilizing physiologic levels of viral gene products expressed from their native promoter(s) in the natural host cells for HPV infection. A quantitative colony-forming assay permits comparison of the capacities of various transfected HPV types and mutant HPV genomes to initially form colonies and immortalize human keratinocytes. In conjunction with additional methods, these protocols enable examination of genomic stability, viral and cellular gene expression, viral integration, and differentiation patterns influenced by HPV persistence in clonal human keratinocytes that effectively mimic early events in HPV infection.

Genome, Viral

Transfection

Keratinocytes - virology

Virus Replication

Feeder Cells

Humans

Cells, Cultured

Fibroblasts

Colony-Forming Units Assay

Papillomaviridae - physiology

Quality Control

Virus Cultivation - methods

Details

Title: Subtitle: Analyzing the Human Papillomavirus (HPV) Life Cycle in Primary Keratinocytes with a Quantitative Colony-Forming Assay
Creators: Michael J Lace - Veterans Healthcare System, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, Iowa; The Department of Pathology, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, Iowa
Lubomir P Turek
James R Anson
Thomas H Haugen
Resource Type: Journal article
Publication Details: Current protocols in microbiology (Online), Vol.33(1), pp.14B.2.1-14B.2.13
Publisher: United States
DOI: 10.1002/9780471729259.mc14b02s33
PMID: 24789595
ISSN: 1934-8525
eISSN: 1934-8533
Language: English
Date published: 05/01/2014
Academic Unit: Pathology
Record Identifier: 9984047601602771

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