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

Jacobsson K, Rosander A, Bjerketorp J, Frykberg L - Biol Proced Online (2003)

Bottom Line: 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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Microbiology, Swedish University of Agricultural Sciences. Box 7025, SE-750 07 UPPSALA. Sweden.

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.

No MeSH data available.


Schematic outline of library construction and panning procedure.
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Figure 1: Schematic outline of library construction and panning procedure.

Mentions: In shotgun phage display, randomly fragmented chromosomal DNA is cloned into a phagemid vector. After transformation into E. coli and infection with helper phage, this results in a library consisting of phages together expressing parts of all proteins encoded by the bacterial genome. By panning against an immobilised ligand, or a mixture of ligands, the gene encoding the corresponding receptin can be identified. The procedure is schematically outlined in Figure 1.


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

Jacobsson K, Rosander A, Bjerketorp J, Frykberg L - Biol Proced Online (2003)

Schematic outline of library construction and panning procedure.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC154567&req=5

Figure 1: Schematic outline of library construction and panning procedure.
Mentions: In shotgun phage display, randomly fragmented chromosomal DNA is cloned into a phagemid vector. After transformation into E. coli and infection with helper phage, this results in a library consisting of phages together expressing parts of all proteins encoded by the bacterial genome. By panning against an immobilised ligand, or a mixture of ligands, the gene encoding the corresponding receptin can be identified. The procedure is schematically outlined in Figure 1.

Bottom Line: 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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Microbiology, Swedish University of Agricultural Sciences. Box 7025, SE-750 07 UPPSALA. Sweden.

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.

No MeSH data available.