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J Virol. 2012 Jan;86(1):594-8. doi: 10.1128/JVI.05988-11. Epub 2011 Oct 19.

Reconstitution of the Kaposi's sarcoma-associated herpesvirus nuclear egress complex and formation of nuclear membrane vesicles by coexpression of ORF67 and ORF69 gene products.

Author information

1
Viral Oncology Program, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Johns Hopkins University, School of Medicine, Baltimore, Maryland, USA. pdesai@jhmi.edu

Abstract

The Kaposi's sarcoma-associated herpesvirus nuclear egress complex is composed of two proteins, ORF67 and ORF69. In this study, we have recapitulated the KSHV complex by coexpression of these two proteins in insect cells using expression from recombinant baculoviruses. The proteins form a complex at the nuclear membrane as judged by live-cell analysis of protein fusions tagged with green fluorescent protein (GFP) and mCherry. Ultrastructural analysis of infected cells showed that ORF67 expression results in reduplication of the nuclear membrane. When the two proteins are expressed together, numerous virion-size nuclear membrane-derived vesicles were evident at the nuclear margins.

PMID:
22013050
PMCID:
PMC3255883
DOI:
10.1128/JVI.05988-11
[Indexed for MEDLINE]
Free PMC Article

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