Immuno-electron microscopy of recombinant SpaFED-piliated lactococcal cells.

Abstract

<p>Immunogold pilin protein-labeling and electron microscopic analysis of recombinant WT (GRS1189) and SpaF pilin-deleted (GRS1126) SpaFED-piliated lactococci (including GRS71 cells as a control) were done using the techniques described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0113922#s4" target="_blank">Materials and Methods</a>. GRS1189 (<b>A and inset</b>) and GRS71 (<b>B</b>) cells are single-labeled with anti-SpaD serum and protein A-10-nm gold particles. GRS1189 cells are double-labeled either with SpaD (10-nm; white arrowhead) and SpaF (15-nm; gray arrowhead) antisera (<b>C</b>) or with SpaD (10-nm; white arrowhead) and SpaE (15-nm; black arrowhead) antisera (<b>D</b>). GRS1226 cells are single-labeled with SpaD antiserum (10-nm) (<b>E</b>) as well as double-labeled either with SpaD (10-nm) and SpaF (15-nm) antisera (<b>F</b>) or with SpaD (10-nm; white arrowhead) and SpaE (15-nm; black arrowhead) antisera (<b>G</b>). A scale bar with dimensions is included in each panel.</p