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Shotgun Phage Display - Selection for Bacterial Receptins or other Exported Proteins

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Author(s): Jacobsson Karin | Rosander Anna | Bjerketorp Joakim | Frykberg Lars

Journal: Biological Procedures Online
ISSN 1480-9222

Volume: 5;
Issue: 1;
Start page: 123;
Date: 2003;
Original page

Keywords: Peptide Library | Staphylococcus aureus

ABSTRACT
Shotgun phage display cloning involves construction of libraries from randomly fragmented bacterial chromosomal DNA, cloned genes, or eukaryotic cDNAs, into a phagemid vector. The library obtained consists of phages expressing polypeptides corresponding to all genes encoded by the organism, or overlapping peptides derived from the cloned gene. From such a library, polypeptides with affinity for another molecule can be isolated by affinity selection, panning. The technique can be used to identify bacterial receptins and identification of their minimal binding domain, and but also to identify epitopes recognised by antibodies. In addition, after modification of the phagemid vector, the technique has also been used to identify bacterial extracytoplasmic proteins.
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