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Generation of a Recombinant Cytomegalovirus for Expression of a Hantavirus Glycoprotein

Albert A. Rizvanov,^1,2 Albert G. M. van Geelen,^1, Sergey Morzunov,^1,3 Elmer W. Otteson,¹ Charlotte Bohlman,^1,2 Gregory S. Pari,^1,2 and Stephen C. St. Jeor^1,2*

Department of Microbiology,¹ Graduate Program in Cell and Molecular Biology, University of Nevada at Reno,² Department of Pathology and Nevada State Health Laboratory, Reno, Nevada 89557³

Received 14 May 2003/ Accepted 29 July 2003

A cytomegalovirus (CMV) was isolated from its natural host, Peromyscus maniculatus, and was designated Peromyscus CMV (PCMV). A recombinant PCMV was constructed that contained Sin Nombre virus glycoprotein G1 (SNV-G1) fused in frame to the enhanced green fluorescent protein (EGFP) gene inserted into a site homologous to the human CMV UL33 (P33) gene. The recombinant CMV was used for expression and immunization of deer mice against SNV-G1. The results of the study indicate that P. maniculatus could be infected with as few as 10 virus particles of recombinant virus. Challenge of P. maniculatus with either recombinant or wild-type PCMV produced no overt pathology in infected animals. P. maniculatus immunized with recombinant virus developed an antibody response to SNV and EGFP. When rechallenged with recombinant virus, animals exhibited an anamnestic response against SNV. Interestingly, a preexisting immune response against PCMV did not prevent reinfection with recombinant PCMV.

Present address: University of California at Davis, Center for Comparative Medicine, Davis, CA 95616.

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