1887

Abstract

strains HB and K28 express surface proteins called Fbe or SdrG, respectively, that have sequence similarity to the clumping factors ClfA and ClfB of . A mutation in the gene of strain HB was isolated by directed plasmid integration using the broad-host-range temperature-sensitive plasmid pGHost9 (pVE6155). An internal fragment of was cloned into pGHost9 and the chimaeric plasmid was mobilized from RN4220 to 9142 by conjugation promoted by plasmid pGO1. The plasmid was then transferred to strain HB by phage-48-mediated transduction. The plasmid integrated into the chromosomal gene at a frequency of 28×10. All the survivors tested had a copy of pGHost9 integrated into the chromosomal gene either as a single copy or as a tandem array. Western immunoblotting showed that the wall-associated Fbe protein was absent in the mutant. Wild-type HB adhered to immobilized fibrinogen in a dose-dependent and saturable fashion whereas the mutant did not bind. The Fbe proteins of HB and K28 were expressed at a high level in MG1363 using the expression vector pKS80. These strains adhered strongly to immobilized fibrinogen. These results confirm that Fbe is a fibrinogen-binding adhesin.

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2001-09-01
2020-01-22
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