Methods for the genetic manipulation of Bornaviruses

Borna disease virus (BDV) has an RNA genome of negative polarity with a total length ca. 8.9 kb. It contains six open reading frames coding for the following proteins: nucleoprotein (N, p40), phosphoprotein (P, p24), matrix protein (M, gp18), surface protein (G, gp92), polymerase (L) and protein of unknown function (X, p10). For the functional analysis of the viral polymerase complex and for the localization of cis-active regulatory elements, we would like to apply "reverse genetics": a novel technology for the genetic manipulation of the viral genome. Such technology has only recently been introduced for work with other negative-stand RNA viruses. In a first step, we try to generate an artificial viral minigenome with large internal deletions that has maintained the ability to replicate after introduction into BDV-infected cells. With the help of this minigenome we will then try to identify the viral proteins that constitue the viral transcriptase and replicase complexes. The ultimate goal of this project is to generate recombinant BDV strains from cloned viral cDNA.

Additional information: http://hhttp://www.ukl.uni-freiburg.de/microbio
contact person: Prof. Dr. Peter Staeheli
Phone: (0761) 203-6579
Email: staeheli@ukl.uni-freiburg.de

Start of project: 1998
End of project: 2003

Albert-Ludwigs-University Freiburg
Stäheli Peter, Schwemmle Martin
Department für Medizinische Mikrobiologie und Hygiene
Institut für Virologie
Hermann-Herder-Strasse 11
79104 Freiburg
Germany

Phone: +49 761 203 6534
Fax: +49 761 203 6626
http://www.virologie.uniklinik-freiburg.de
Actual Research Report

Bornaviren

• Pleschka St, Staeheli P, Kolodziejek J, Richt JA, Nowotny N, Schwemmle M.: Conservation of coding potential and terminal sequences in four different isolates of Borna disease virus. J Gen Virol, 2001; 82: 2681-2690.
• Staeheli P, Sentandreu M, Pagenstecher A, Hausmann J.: Alpha/beta Interferon promotes transcription and inhibits replication of Borna Disease Virus in persistently infected cells. J Virol, 2001; 75: 8216-8223.