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Research article2002Peer reviewed

Phage display as a novel screening method to identify extracellular proteins

Rosander, A; Bjerketorp, J; Frykberg, L; Jacobsson, K

Abstract

Extracellular proteins are involved in many diverse and essential cell functions and in pathogenic bacteria, and they may also serve as virulence factors. Therefore, there is a need for methods that identify the genes encoding this group of proteins in a bacterial genome. Here, we present such a method based on the phage display technology. A novel gene III-based phagemid vector, pG3DSS, was constructed that lacks the signal sequence which normally orientates the encoded fusion protein to the Escherichia coli cell membrane, where it is assembled into the phage particle. When randomly fragmented DNA is inserted into this vector, only phagemids containing an insert encoding a signal sequence will give rise to phage particles displaying a fusion protein. These phages also display an E-tag epitope in fusion with protein 111, which enables isolation of phages displaying a fusion protein, using antibodies against the epitope. From a library constructed from Staphylococcus aureus chromosomal DNA, genes encoding secreted as well as transmembrane proteins were isolated, including adhesins, enzymes and transport proteins. (C) 2002 Elsevier Science B.V. All rights reserved.

Keywords

extracellular proteins; phage display; protein export; signal sequence; Staphylococcus aureus

Published in

Journal of Microbiological Methods
2002, volume: 51, number: 1, article number: PII S0167-7012(02)00052-0

SLU Authors

UKÄ Subject classification

Microbiology
Biochemistry and Molecular Biology

Publication identifier

  • DOI: https://doi.org/10.1016/S0167-7012(02)00052-0

Permanent link to this page (URI)

https://res.slu.se/id/publ/90881