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A morphologic study of filamentous phage infection of Escherichia coli using biotinylated phages
Author(s) -
Nakamura Michihiro,
Tsumoto Kouhei,
Kumagai Izumi,
Ishimura Kazunori
Publication year - 2003
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(03)00050-4
Subject(s) - periplasmic space , bacterial outer membrane , biotinylation , staining , inner membrane , escherichia coli , biology , negative stain , microbiology and biotechnology , membrane , biochemistry , electron microscope , genetics , physics , optics , gene
Using biotinylated phage (BIO‐phages), we observed the infection of filamentous phages into Escherichia coli JM109 morphologically. BIO‐phages and BIO‐phage‐derived proteins, mainly pVIII, were detected in E. coli by using the avidin–biotin–peroxidase complex method with electron microscopy. Infected cells revealed positive staining on the outer and inner membranes and in the periplasmic space. Some cells showed specific or predominant staining of the outer membrane, whereas others showed predominant staining of the inner membrane or equivalent staining of the outer and inner membranes. The periplasmic spaces in some infected cells were expanded and filled with reaction products. Some cells showed wavy lines of positive staining in the periplasmic space. BIO‐phages were detected as thick filaments or clusters covered with reaction products. The ends of the infecting phages were located on the surface of cells, in the periplasmic space, or on the inner membrane. These findings suggest that phage major coat proteins are integrated into the outer membrane and that phages cause periplasmic expansion during infection.