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Journal of Virology, October 2001, p. 9282-9286, Vol. 75, No. 19
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.19.9282-9286.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Rescue of Recombinant Thogoto Virus from Cloned cDNA

Elke Wagner, Othmar G. Engelhardt, Simone Gruber, Otto Haller, and Georg Kochs*

Abteilung Virologie, Institut für Medizinische Mikrobiologie und Hygiene, Universität Freiburg, D-79008 Freiburg, Germany

Received 11 April 2001/Accepted 21 June 2001

Thogoto virus (THOV) is a tick-transmitted orthomyxovirus with a genome consisting of six negative-stranded RNA segments. To rescue a recombinant THOV, the viral structural proteins were produced from expression plasmids by means of a vaccinia virus expressing the T7 RNA polymerase. Genomic virus RNAs (vRNAs) were generated from plasmids under the control of the RNA polymerase I promoter. Using this system, we could efficiently recover recombinant THOV following transfection of 12 plasmids into 293T cells. To verify the recombinant nature of the rescued virus, specific genetic tags were introduced into two vRNA segments. The availability of this efficient reverse genetics system will allow us to address hitherto-unanswered questions regarding the biology of THOV by manipulating viral genes in the context of infectious virus.

* Corresponding author. Mailing address: Abteilung Virologie, Institut für Medizinische Mikrobiologie und Hygiene, Universität Freiburg, D-79008 Freiburg, Germany. Phone: 49-761-2036623. Fax: 49-761-2036562. E-mail: KOCHS{at}UKL.UNI-FREIBURG.DE.

Journal of Virology, October 2001, p. 9282-9286, Vol. 75, No. 19
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.19.9282-9286.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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