# Core C - Virus Core

> **NIH NIH P01** · UNIVERSITY OF WISCONSIN-MADISON · 2022 · $78,528

## Abstract

CORE C – PROJECT SUMMARY/ABSTRACT
The purpose of Core C (Virus Core) is to provide a time and cost efficient mechanism for the generation,
validation, distribution, and archival storage of wild-type (WT) and mutant viruses needed by all 8 research
groups associated with this Program Project for their proposed studies. The Specific Aims are: (1) to produce
validated stocks of WT, mutant, and revertant Epstein-Barr virus (EBV) and Kaposi Sarcoma Herpesvirus
(KSHV); (2) to produce validated stocks of WT and mutant hepatitis B viruses (HBVs); (3) to produce validated
stocks of infectious mouse papillomavirus (MmuPV1) and human papillomavirus (HPV) pseudoviruses.
(1).
Mutant EBV (for Projects 3, 4, and 5) or KSHV (for Project 3) will be generated in E. coli starting with
appropriate BACs using the scarless En Passant method In addition to standard methods, we have
established a bioinformatic pipeline to analyze next generation sequencing data of herpesvirus genomes to
ensure there are no unintended mutations within the unique regions. Whenever necessary, phenotypes will
be validated using transcomplementation or the construction of revertant EBV/KSHV genomes. Virus stocks
of WT, mutant, and revertant variants of EBV or KSHV will be generated using a protocol developed by Dr.
Sugden, titered, and stored for use by all four EBV groups. HBV virions will be produced (for Project 2) from
HepAD38 cells, a HepG2 derivative that is stably transfected with a tetracycline-inducible HBV expression
system. HBV mutants will be constructed by recombinant DNA approaches in vectors already developed by
the Loeb laboratory. Papillomavirus virions and pseudovirions will be generated (for Project 1) by co-
transfection of 293T cells with (i) the desired capsid protein-expression plasmids, and (ii) the desired viral or
luciferase/GFP-encoding DNAs targeted for encapsidation using protocols previously published by the
Lambert/Ahlquist laboratories. In addition to increased cost efficiency and quality control, the existence of
this Core facilitates the use of common virus stocks that will make the interpretation of complementary data
generated among the various research groups more reliably merged toward achieving shared aims within the
projects. All 8 research groups associated with the 5 projects will be served by this Core facility as needed.

## Key facts

- **NIH application ID:** 10414882
- **Project number:** 5P01CA022443-45
- **Recipient organization:** UNIVERSITY OF WISCONSIN-MADISON
- **Principal Investigator:** ERIC C. JOHANNSEN
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $78,528
- **Award type:** 5
- **Project period:** 1997-02-01 → 2024-02-29

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10414882

## Citation

> US National Institutes of Health, RePORTER application 10414882, Core C - Virus Core (5P01CA022443-45). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10414882. Licensed CC0.

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