title: Development of an improved reverse genetics system for Akabane bunyavirus.
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.
authors.affiliation: Department of Veterinary Microbiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan; Department of Infection Control and Disease Prevention, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. Electronic address: atakiko@mail.ecc.u-tokyo.ac.jp.
authors.forename: Akiko
authors.initials: A
authors.lastname: Takenaka-Uema
authors.affiliation: Department of Veterinary Microbiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. Electronic address: sugiura_keita@kaken.co.jp.
authors.forename: Keita
authors.initials: K
authors.lastname: Sugiura
authors.affiliation: Department of Veterinary Microbiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan; Department of Preclinical and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, 999 Phutthamonthon Sai 4 Road Salaya, Phutthamonthon Nakhonpathom 73170, Thailand. Electronic address: norasuthi.ban@mahidol.ac.th.
authors.forename: Norasuthi
authors.initials: N
authors.lastname: Bangphoomi
authors.affiliation: Department of Veterinary Pathology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. Electronic address: chieko_shioda@nm.maff.go.jp.
authors.forename: Chieko
authors.initials: C
authors.lastname: Shioda
authors.affiliation: Department of Veterinary Pathology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. Electronic address: auchidak@mail.ecc.u-tokyo.ac.jp.
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Uchida
authors.affiliation: Department of Veterinary Microbiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan; Research Unit for Global Infection Control, National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan. Electronic address: kkato@obihiro.ac.jp.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Kato
authors.affiliation: Department of Infection Control and Disease Prevention, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. Electronic address: ahaga@mail.ecc.u-tokyo.ac.jp.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Haga
authors.affiliation: Department of Veterinary Microbiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. Electronic address: amurakam@mail.ecc.u-tokyo.ac.jp.
authors.forename: Shin
authors.initials: S
authors.lastname: Murakami
authors.affiliation: Department of Veterinary Microbiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. Electronic address: akashih@mail.ecc.u-tokyo.ac.jp.
authors.forename: Hiroomi
authors.initials: H
authors.lastname: Akashi
authors.affiliation: Department of Veterinary Microbiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. Electronic address: ahorimo@mail.ecc.u-tokyo.ac.jp.
authors.forename: Taisuke
authors.initials: T
authors.lastname: Horimoto
mesh_terms: D000818:Animals; D002044:Bunyaviridae Infections / Q000662:veterinary / Q000821:virology; D002460:Cell Line; D007564:Japan; D009154:Mutation; D016851:Orthobunyavirus / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity / Q000502:physiology*; D059386:Reverse Genetics / Q000379:methods*; D037521:Virulence Factors / Q000235:genetics
chemical_list: D037521:Virulence Factors
keywords: Akabane virus; Reverse genetics; Vaccine