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Elsevier, Virology, 2(305), p. 406-414, 2003

DOI: 10.1006/viro.2002.1748

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Isolation and Characterisation of the Rabies Virus N°-P Complex Produced in Insect Cells

This paper is made freely available by the publisher.
This paper is made freely available by the publisher.

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Abstract

When the nucleoprotein (N) of nonsegmented negative-strand RNA viruses is expressed in insect cells, it binds to cellular RNA and forms N-RNA complexes just like viral nucleocapsids. However, in virus-infected cells, N is prevented from binding to cellular RNA because a soluble complex is formed between N and the viral phosphoprotein (P), the N degrees -P complex. N is only released from this complex for binding to newly made viral or complementary RNA. We coexpressed rabies virus N and P proteins in insect cells and purified the N degrees -P complex. Characterisation by gel filtration, polyacrylamide gel electrophoresis, analytical ultracentrifugation, native mass spectroscopy, and electron microscopy showed that the complex consists of one N protein plus two P proteins, i.e., an N degrees -P(2) complex.