Construction and characterization of a fluorescent sendai virus carrying the gene for envelope fusion protein fused with enhanced green fluorescent protein.
Sendai virus (SeV) is an enveloped virus with a non-segmented negative-strand RNA genome. SeV envelope fusion (F) glycoproteins play crucial roles in the viral life cycle in processes such as viral binding, assembly, and budding. In this study, we developed a viable recombinant SeV designated F-EGFP SeV/ΔF, in which the F protein was replaced by an F protein fused to EGFP at the carboxyl terminus. Living infected cells of the recombinant virus were directly visualized by green fluorescence. The addition of EGFP to the F protein maintained the activities of the F protein in terms of intracellular transport to the plasma membrane via the ER and the Golgi apparatus and fusion activity in the infected cells. These results suggest that this fluorescent SeV is a useful tool for studying the viral binding, assembly, and budding mechanisms of F proteins and the SeV life cycle in living infected cells.
Department of Agro-Bioscience, Faculty of Agriculture, Iwate University, Morioka, Japan.
Masao
M
Miyazaki
Hiroaki
H
Segawa
Tetsuro
T
Yamashita
Yafeng
Y
Zhu
Kaoru
K
Takizawa
Mamoru
M
Hasegawa
Hideharu
H
Taira
D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000235:genetics*; D021381:Protein Transport; D011993:Recombinant Fusion Proteins; D012107:Research Design; D029082:Sendai virus / Q000235:genetics*; D014759:Viral Envelope Proteins / Q000235:genetics*; D014760:Viral Fusion Proteins / Q000502:physiology
D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins