A visualization method of filamentous phage infection and phage-derived proteins in Escherichia coli using biotinylated phages.
Direct visualization of filamentous phage infection in Escherichia coli (E. coli) was attempted using biotinylated phages (BIO-phages). The biotinylation of the phages did not influence their infectivity into E. coli. E. coli infected with BIO-phages could be detected by using fluorescein-conjugated avidin with confocal laser scanning microscopy, and BIO-phages and BIO-phage-derived proteins in E. coli could be directly observed by using the avidin-biotin-peroxidase complex method with electron microscopy. This is the first report of direct visualization of phage infection and phage-derived proteins in the host cell using a biotin-avidin interaction. This simple and powerful method is applicable to the study of infection by various viruses.
Nakamura
M
M
Department of Anatomy and Cell Biology, School of Medicine, Tokushima University, 3-18-15 Kuramoto, Tokushima 770-8503, Japan.
Tsumoto
K
K
Ishimura
K
K
Kumagai
I
I
D001360:Avidin; D001710:Biotin; D004926:Escherichia coli / Q000648:ultrastructure / Q000821:virology*; D017912:Inovirus / Q000472:pathogenicity / Q000502:physiology* / Q000648:ultrastructure; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D014764:Viral Proteins / Q000378:metabolism
D014764:Viral Proteins; D001360:Avidin; D001710:Biotin