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Journal of Virology, March 2007, p. 2957-2969, Vol. 81, No. 6
0022-538X/07/$08.00+0     doi:10.1128/JVI.01997-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Construction of an Infectious Rhesus Rhadinovirus Bacterial Artificial Chromosome for the Analysis of Kaposi's Sarcoma-Associated Herpesvirus-Related Disease Development{triangledown}

Ryan D. Estep,1 Michael F. Powers,1 Bonnie K. Yen,1 He Li,1 and Scott W. Wong1,2,3*

Vaccine and Gene Therapy Institute, Oregon Health & Science University West Campus,1 Division of Pathobiology and Immunology, Oregon National Primate Research Center, Beaverton, Oregon 97006,2 Department of Molecular Microbiology and Immunology, Oregon Health & Science University, Portland, Oregon 972013

Received 13 September 2006/ Accepted 28 December 2006

Rhesus rhadinovirus (RRV) is closely related to Kaposi's sarcoma-associated herpesvirus (KSHV)/human herpesvirus 8 (HHV-8) and causes KSHV-like diseases in immunocompromised rhesus macaques (RM) that resemble KSHV-associated diseases including multicentric Castleman's disease and non-Hodgkin's lymphoma. RRV retains a majority of open reading frames (ORFs) postulated to be involved in the pathogenesis of KSHV and is the closest available animal model to KSHV infection in humans. Here we describe the generation of a recombinant clone of RRV strain 17577 (RRV17577) utilizing bacterial artificial chromosome (BAC) technology. Characterization of the RRV BAC demonstrated that it is a pathogenic molecular clone of RRV17577, producing virus that behaves like wild-type RRV both in vitro and in vivo. Specifically, BAC-derived RRV displays wild-type growth properties in vitro and readily infects simian immunodeficiency virus-infected RM, inducing B cell hyperplasia, persistent lymphadenopathy, and persistent infection in these animals. This RRV BAC will allow for rapid genetic manipulation of the RRV genome, facilitating the creation of recombinant versions of RRV that harbor specific alterations and/or deletions of viral ORFs. This system will provide insights into the roles of specific RRV genes in various aspects of the viral life cycle and the RRV-associated pathogenesis in vivo in an RM model of infection. Furthermore, the generation of chimeric versions of RRV containing KSHV genes will allow analysis of the function and contributions of KSHV genes to viral pathogenesis by using a relevant primate model system.

* Corresponding author. Mailing address: Vaccine and Gene Therapy Institute, Oregon Health & Science University, West Campus, 505 N.W. 185th Avenue, Beaverton, OR 97006. Phone: (503) 690-5285. Fax: (503) 418-2719. E-mail: wongs{at}ohsu.edu.

{triangledown} Published ahead of print on 10 January 2007.

Journal of Virology, March 2007, p. 2957-2969, Vol. 81, No. 6
0022-538X/07/$08.00+0     doi:10.1128/JVI.01997-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Zhou, F., Li, Q., Gao, S.-J. (2009). A sequence-independent in vitro transposon-based strategy for efficient cloning of genomes of large DNA viruses as bacterial artificial chromosomes. Nucleic Acids Res 37: e2-e2 [Abstract] [Full Text]  
  • Orzechowska, B. U., Powers, M. F., Sprague, J., Li, H., Yen, B., Searles, R. P., Axthelm, M. K., Wong, S. W. (2008). Rhesus macaque rhadinovirus-associated non-Hodgkin lymphoma: animal model for KSHV-associated malignancies. Blood 112: 4227-4234 [Abstract] [Full Text]  


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