Use of the red fluorescent protein as a marker of Kaposi's sarcoma-associated herpesvirus lytic gene expression

Virology
Jeffrey Vieira, Patricia M O'Hearn

Abstract

A hallmark of all herpesvirus is the ability to exist in either a latent, or lytic, state of replication, enabling the lifelong infection of its host. Kaposi's sarcoma (KS)-associated herpesvirus (KSHV) can efficiently establish a latent infection in a variety of cell types in vitro, making it a valuable model for the study of latency and reactivation. To facilitate the identification of KSHV lytic replication, and allow subsequent experiments with live cells, a recombinant virus, rKSHV.219, was constructed using JSC-1 cells that expresses the red fluorescent protein (RFP) from the KSHV lytic PAN promoter, the green fluorescent protein (GFP) from the EF-1alpha promoter, and with the gene for puromycin resistance as a selectable marker. rKSHV.219 from JSC-1 cells was used to infect Vero cells for purification of the recombinant virus. Vero cells were also used for the production of rKSHV.219 at levels of 10(5)-10(6) infectious units (IU) of virus per milliliter using a combination of KSHV/RTA expressed from a baculovirus vector, BacK50, and butyrate. Virus produced from Vero cells was used to infect human fibroblasts (HF), 293, DU145, T24, HaCaT, and HEp-2 cells, and in all cells except 293 cells, only a latent infection was est...Continue Reading

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Citations

Feb 12, 2014·Proceedings of the National Academy of Sciences of the United States of America·Lin-Xu WangCharles Wood
Jun 4, 2015·Emerging Microbes & Infections·Masahiko Ajiro, Zhi-Ming Zheng
Feb 19, 2013·Cell Host & Microbe·Patrick J DillonBlossom Damania
Oct 29, 2005·Journal of Virology·Lei Chen, Michael Lagunoff
Oct 20, 2006·Journal of Virology·Melanie M BrinkmannThomas F Schulz
Mar 17, 2007·The Journal of Clinical Investigation·Grzegorz SarekPäivi M Ojala
Dec 14, 2007·Journal of Virology·Gaby SanderMichael Stürzl
Jul 31, 2009·Journal of Virology·Carlos M GonzálezBlossom Damania
Oct 3, 2009·PLoS Pathogens·Heesoon ChangJae U Jung
Mar 25, 2010·PLoS Pathogens·Grzegorz SarekPäivi M Ojala
Apr 7, 2010·The Journal of Clinical Investigation·Don Ganem
Jul 28, 2010·PLoS Pathogens·Zsolt TothJae U Jung
Oct 29, 2010·Journal of Virology·John A WestBlossom Damania
Mar 16, 2011·Antimicrobial Agents and Chemotherapy·Soren GanttJeffrey Vieira

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