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Construction and Characterization of a Fluorescent Sendai Virus Carrying the Gene for Envelope Fusion Protein Fused with Enhanced Green Fluorescent Protein
Author(s) -
Masao Miyazaki,
Hiroaki Segawa,
Tetsuro Yamashita,
Yafeng Zhu,
Kaoru Takizawa,
Mamoru Hasegawa,
Hideharu Taira
Publication year - 2010
Publication title -
bioscience biotechnology and biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.509
H-Index - 116
eISSN - 1347-6947
pISSN - 0916-8451
DOI - 10.1271/bbb.100511
Subject(s) - sendai virus , green fluorescent protein , viral envelope , fusion protein , biology , glycoprotein , recombinant dna , virus , viral life cycle , microbiology and biotechnology , budding , transfection , fusion gene , golgi apparatus , virology , gene , biochemistry , viral replication , endoplasmic reticulum
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.

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