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Updated: Jul 6, 2026

Electrophoretic Analysis of Replication Through Structure-Prone DNA Repeats Within the SV40-Based Human Episome

Electrophoretic Analysis of Replication Through Structure-Prone DNA Repeats Within the SV40-Based Human Episome

Published on: September 13, 2024

SV40 reporter viruses.

Rebecca B Katzman¹, Mark Seeger, Kathleen Rundell

¹Department of Microbiology-Immunology and The Robert H. Lurie Comprehensive Cancer Center, Northwestern University, 303 E. Chicago Avenue, Chicago, IL 60611, USA. rebeccab@northwestern.edu

Journal of Virological Methods

|April 12, 2008

Summary

This summary is machine-generated.

Researchers developed three simian virus 40 (SV40) reporter viruses. One virus, expressing green fluorescent protein (GFP), aids live-cell imaging, while tagged variants help detect SV40 infections in cells with existing large-T antigen.

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Electrophoretic Analysis of Replication Through Structure-Prone DNA Repeats Within the SV40-Based Human Episome

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Area of Science:

Virology
Molecular Biology
Cell Biology

Background:

Simian virus 40 (SV40) is a polyomavirus with significant implications in research and oncology.
Understanding SV40 replication and infection dynamics is crucial for developing antiviral strategies and cancer therapies.
Reporter viruses are essential tools for tracking viral infection and gene expression in real-time.

Purpose of the Study:

To construct and characterize novel SV40 reporter viruses for enhanced infection monitoring.
To evaluate the utility of green fluorescent protein (GFP) and FLAG epitope-tagged SV40 constructs.
To assess the efficiency of reporter virus replication and particle production.

Main Methods:

Construction of three distinct SV40 reporter viruses: one with a GFP fusion protein, and two with FLAG epitope tags at the N- and C-termini of the large-T antigen.
Titration of reporter viruses to determine infectious particle production and compare with wild-type (WT) SV40.
Assessment of reporter virus functionality in detecting superinfecting SV40 in cells with pre-existing nuclear large-T antigen.

Main Results:

A GFP-expressing SV40 reporter virus was created for live-cell imaging applications.
The vC-LT(FLAG) reporter virus demonstrated high titers, comparable to WT SV40, indicating efficient replication and spread.
The vN-LT(FLAG) construct, while viable, exhibited defects in infectious particle production and spread.
All constructed reporter viruses proved effective in detecting superinfecting SV40 in cells already expressing nuclear large-T antigen.

Conclusions:

Novel SV40 reporter viruses, including a GFP-based system for live imaging and FLAG-tagged variants, have been successfully developed.
The vC-LT(FLAG) construct serves as a robust reporter for SV40 infection studies due to its high viral titers.
These reporter viruses offer valuable tools for studying SV40 infection dynamics, particularly in complex cellular environments like persistently infected cells.