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Cloning and Expression of the Gene for a Fibronectin-Binding Protein From Staphylococcus Aureus

Cloning and Expression of the Gene for a Fibronectin-Binding Protein From Staphylococcus Aureus

J I Flock, G Fröman, K Jönsson, B Guss, C Signäs, B Nilsson, G Raucci, M Höök, T Wadström, M Lindberg

EMBO J. 1987 Aug;6(8):2351-7.

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42411046	Protein A (extracellular)-Agarose from Staphylococcus aureus			Price

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