Journal article

Authors list: Blakqori, G; Kochs, G; Haller, O; Weber, F

Publication year: 2003

Pages: 1207-1214

Journal: Journal of General Virology

Volume number: 84

Issue number: 5

ISSN: 0022-1317

DOI Link: https://doi.org/10.1099/vir.0.18876-0

Publisher: Microbiology Society

Abstract: 

La Crosse virus (LACV), a member of the family Bunyaviridae, is the primary cause of paediatric; encephalitis in the United States. In this study, a functional RNA polymerase Q gene of LACV was cloned and a reverse genetics system established. A reporter minireplicon mimicking the viral genome was constructed by flanking the Renilla luciferase gene with the 3' and 5' noncoding regions of the genomic M segment. These noncoding regions serve as promoters for the viral polymerase. Both L and nucleocapsid (N) genes were expressed by means of T7 RNA polymerase, which was provided by the recombinant T7-expressing modified vaccinia virus Ankara. Renilla reporter activity in transfected cells reflected reconstitution of recombinant nucleocapsids by functional L and N gene products. Time-course experiments revealed a rapid increase in minireplicon activity from 10 to 18 h after the onset of L and N expression. Minireplicon activity was found to be dependent on the correct ratio of L to N plasmids, with too much of either construct resulting in downregulation. Furthermore, a specific inhibitory effect of LACV NSS protein on minireplicon activity was found. In passaging experiments using parental helper virions, it was demonstrated that the recombinant nucleocapsids are a useful model for transcription, replication and packaging of LACV.

Harvard Citation style: Blakqori, G., Kochs, G., Haller, O. and Weber, F. (2003) Functional L polymerase of La Crosse virus allows in vivo reconstitution of recombinant nucleocapsids, Journal of General Virology, 84(5), pp. 1207-1214. https://doi.org/10.1099/vir.0.18876-0

APA Citation style: Blakqori, G., Kochs, G., Haller, O., & Weber, F. (2003). Functional L polymerase of La Crosse virus allows in vivo reconstitution of recombinant nucleocapsids. Journal of General Virology. 84(5), 1207-1214. https://doi.org/10.1099/vir.0.18876-0