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. 1987 Aug;6(8):2351–2357. doi: 10.1002/j.1460-2075.1987.tb02511.x

Cloning and expression of the gene for a fibronectin-binding protein from Staphylococcus aureus.

J I Flock 1, G Fröman 1, K Jönsson 1, B Guss 1, C Signäs 1, B Nilsson 1, G Raucci 1, M Höök 1, T Wadström 1, M Lindberg 1
PMCID: PMC553639  PMID: 2822388

Abstract

The gene encoding the fibronectin-binding protein (FNBP) from Staphylococcus aureus strain 8325-4 was isolated from a gene bank in pBR322. The original clone, containing a 6.5-kb insert, gave a functional product present in the periplasm of Escherichia coli. Analysis of polypeptides isolated after affinity chromatography on fibronectin-Sepharose followed by ion-exchange chromatography revealed two gene products, 87 and 165 kd in mol. wt. The amino acid compositions of these two polypeptides and a native FNBP from S. aureus strain Newman were very similar. Antibodies raised against the native FNBP from strain Newman precipitated the 125I-labelled 165-kd polypeptide, and unlabeled 165- and 87-kd polypeptides as well as native FNBP inhibited the immunoprecipitation reactions. The region of the fnbp-gene encoding the fibronectin-binding activity has been identified and subcloned in an expression vector based on the staphylococcal protein A gene. The resulting product in E. coli is an extracellular fusion protein consisting of two IgG-binding domains of protein A followed by a fibronectin-binding region. The fusion protein binds to fibronectin and completely inhibits the binding of fibronectin to intact cells of S. aureus.

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Selected References

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