TY - JOUR
T1 - Development of an improved reverse genetics system for Akabane bunyavirus
AU - Takenaka-Uema, Akiko
AU - Sugiura, Keita
AU - Bangphoomi, Norasuthi
AU - Shioda, Chieko
AU - Uchida, Kazuyuki
AU - Kato, Kentaro
AU - Haga, Takeshi
AU - Murakami, Shin
AU - Akashi, Hiroomi
AU - Horimoto, Taisuke
N1 - Funding Information:
We thank Dr. Ito of the Laboratory of Zoonotic Diseases, Gifu University, Japan, for providing BHK/T7-9 cells and Dr. Weber of the Department of Virology, University of Freiburg, Freiburg, Germany, for providing pT7riboSM2. This study was supported in part by a Research and Development Project for Application in Promoting New Policies in Agriculture, Forestry and Fisheries grant from the Ministry of Agriculture, Forestry and Fisheries , and a Grant-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology of Japan .
Publisher Copyright:
© 2016 Elsevier B.V.
PY - 2016/6/1
Y1 - 2016/6/1
N2 - 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.
AB - 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.
KW - Akabane virus
KW - Reverse genetics
KW - Vaccine
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U2 - 10.1016/j.jviromet.2015.12.014
DO - 10.1016/j.jviromet.2015.12.014
M3 - Article
C2 - 26927704
AN - SCOPUS:84960153721
SN - 0166-0934
VL - 232
SP - 16
EP - 20
JO - Journal of Virological Methods
JF - Journal of Virological Methods
ER -
