Production of novel ebola virus-like particles from cDNAs: an alternative to ebola virus generation by reverse genetics.
We established a plasmid-based system for generating infectious Ebola virus-like particles (VLPs), which contain an Ebola virus-like minigenome consisting of a negative-sense copy of the green fluorescent protein gene. This system produced nearly 10(3) infectious particles per ml of supernatant, equivalent to the titer of Ebola virus generated by a reverse genetics system. Interestingly, infectious Ebola VLPs were generated, even without expression of VP24. Transmission and scanning electron microscopic analyses showed that the morphology of the Ebola VLPs was indistinguishable from that of authentic Ebola virus. Thus, this system allows us to study Ebola virus entry, replication, and assembly without biosafety level 4 containment. Furthermore, it may be useful in vaccine production against this highly pathogenic agent.
Watanabe
Shinji
S
Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
Watanabe
Tokiko
T
Noda
Takeshi
T
Takada
Ayato
A
Feldmann
Heinz
H
Jasenosky
Luke D
LD
Kawaoka
Yoshihiro
Y
D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D018076:DNA, Complementary / Q000235:genetics*; D029043:Ebolavirus / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D010957:Plasmids / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism; D014771:Virion / Q000378:metabolism*; D014773:Virology / Q000379:methods
D018076:DNA, Complementary; D008164:Luminescent Proteins; D012367:RNA, Viral; D014759:Viral Envelope Proteins; D049452:Green Fluorescent Proteins