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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.


Related in: MedlinePlus

A filter paper is placed in a glass Petri dish and a few ml of chloroform poured onto the filter paper. The Nitrocellulose-filter, with colonies facing up, is placed on top of some small inert objects (like caps or small metal balls), which will keep the filter from touching the liquid chloroform. After 10 minutes in the closed chamber, the colonies are lysed.
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Figure 4: A filter paper is placed in a glass Petri dish and a few ml of chloroform poured onto the filter paper. The Nitrocellulose-filter, with colonies facing up, is placed on top of some small inert objects (like caps or small metal balls), which will keep the filter from touching the liquid chloroform. After 10 minutes in the closed chamber, the colonies are lysed.

Mentions: Transfer the colonies to a nitrocellulose filter and lyse the cells in chloroform vapour. See Figure 4.


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

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

A filter paper is placed in a glass Petri dish and a few ml of chloroform poured onto the filter paper. The Nitrocellulose-filter, with colonies facing up, is placed on top of some small inert objects (like caps or small metal balls), which will keep the filter from touching the liquid chloroform. After 10 minutes in the closed chamber, the colonies are lysed.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC154567&req=5

Figure 4: A filter paper is placed in a glass Petri dish and a few ml of chloroform poured onto the filter paper. The Nitrocellulose-filter, with colonies facing up, is placed on top of some small inert objects (like caps or small metal balls), which will keep the filter from touching the liquid chloroform. After 10 minutes in the closed chamber, the colonies are lysed.
Mentions: Transfer the colonies to a nitrocellulose filter and lyse the cells in chloroform vapour. See Figure 4.

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.


Related in: MedlinePlus