Development of an improved reverse genetics system for Akabane bunyavirus

Akiko Takenaka-Uema, Keita Sugiura, Norasuthi Bangphoomi, Chieko Shioda, Kazuyuki Uchida, Kentaro Kato, Takeshi Haga, Shin Murakami, Hiroomi Akashi, Taisuke Horimoto

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Akabane disease, caused by the insect-transmitted Akabane virus (AKAV), affects livestock by causing life-threatening deformities or mortality of fetuses. Therefore, Akabane disease has led to notable economic losses in numerous countries, including Japan. In this short communication, a new T7 RNA polymerase-based AKAV reverse genetics system was developed. Using this system, in which three plasmids transcribing antigenomic RNAs were transfected into cells stably expressing T7 polymerase, we successfully reconstituted the live attenuated vaccine TS-C2 strain (named rTTT), and also generated a mutant AKAV (rTTTδNSs) that lacked the gene encoding the nonstructural NSs protein, which is regarded as a virulence factor. Analysis of growth kinetics revealed that rTTTδNSs grew at a much slower rate than the rTTT and TS-C2 virus. These results suggest that our established reverse genetics system is a powerful tool that can be used for AKAV vaccine studies with gene-manipulated viruses.

Original languageEnglish
Pages (from-to)16-20
Number of pages5
JournalJournal of Virological Methods
Volume232
DOIs
Publication statusPublished - 2016 Jun 1
Externally publishedYes

Keywords

  • Akabane virus
  • Reverse genetics
  • Vaccine

ASJC Scopus subject areas

  • Virology

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  • Cite this

    Takenaka-Uema, A., Sugiura, K., Bangphoomi, N., Shioda, C., Uchida, K., Kato, K., Haga, T., Murakami, S., Akashi, H., & Horimoto, T. (2016). Development of an improved reverse genetics system for Akabane bunyavirus. Journal of Virological Methods, 232, 16-20. https://doi.org/10.1016/j.jviromet.2015.12.014