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J Gen Virol 84 (2003), 3099-3104; DOI 10.1099/vir.0.19467-0

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© 2003 Society for General Microbiology

Short Communication

A reverse genetics system for Borna disease virus

Mar Perez, Ana Sanchez, Beatrice Cubitt, Debralee Rosario and Juan Carlos de la Torre

The Scripps Research Institute, Department of Neuropharmacology IMM-6, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA

Correspondence
Juan Carlos de la Torre
juanct{at}scripps.edu

Borna disease virus (BDV) is an enveloped virus. Its non-segmented, negative-stranded RNA genome has the coding capability for six main polypeptides and has an organization characteristic of members of the order Mononegavirales. However, based on its unique genetics and biological features, BDV is considered to be the prototypic member of a new virus family, Bornaviridae. Here, the establishment of a reverse genetics system for BDV is described. Intracellular synthesis of a BDV RNA analogue or minigenome (MG) from a plasmid was driven by RNA polymerase I. Co-transfection with plasmids expressing the BDV polymerase (L), nucleoprotein (N) and phosphoprotein (P) under the control of RNA polymerase II allowed for BDV MG replication and expression. This process depended on a delicate N : P ratio, whereas the L : P ratio was less critical. Two isoforms of N, Np40 and Np38, are present in BDV-infected cells but only Np40 was strictly required for virus polymerase activity. BDV p10 polypeptide encoded by the P gene exhibited a strong inhibitory effect on BDV MG expression.

Published ahead of print on 6 August 2003 as DOI 10.1099/vir.0.19467-0.




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