Gene VIII-Based, Phage-Display Vectors for Selection Against Complex Mixtures of Ligands
Selection of shotgun phage-display libraries against complex mixtures of components, such as cells or sera, may result in a high number of nonspecifically binding phage. Consequently, correct interactions may be difficult to identify. To enable discrimination between faithful and nonspecific interac...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
1998-02-01
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| Series: | BioTechniques |
| Online Access: | https://www.future-science.com/doi/10.2144/98242rr01 |
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| _version_ | 1850153744113598464 |
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| author | Karin Jacobsson Lars Frykberg |
| author_facet | Karin Jacobsson Lars Frykberg |
| author_sort | Karin Jacobsson |
| collection | DOAJ |
| description | Selection of shotgun phage-display libraries against complex mixtures of components, such as cells or sera, may result in a high number of nonspecifically binding phage. Consequently, correct interactions may be difficult to identify. To enable discrimination between faithful and nonspecific interactions, a set of eight different gene VIII-based, phage-display vectors were constructed. All vectors contain a “universal” screening tag positioned in such a way that it is only expressed when the inserted DNA encodes an open reading frame, which corrects a shift of reading frames in the vector. A Staphylococcus aureus shotgun phagedisplay library was made in a stoichiometric mixture of all vectors. After affinity-selection against IgG, one vector completely outcompeted the others. This vector contains the promoter and signal sequence from the gene encoding staphylococcal protein A and one suppressible stop codon immediately upstream of gene VIII. An increase in the frequency of clones expressing the affinity tag in all pannings correlated with selection for ligand-binding clones. This enables detection of putatively correct clones after selection of a shotgun phagedisplay library both against purified ligands and more complex materials like calf serum. |
| format | Article |
| id | doaj-art-ee9ab029c99249fcadf1fbbd40c3aa56 |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 1998-02-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-ee9ab029c99249fcadf1fbbd40c3aa562025-08-20T02:25:37ZengTaylor & Francis GroupBioTechniques0736-62051940-98181998-02-0124229430110.2144/98242rr01Gene VIII-Based, Phage-Display Vectors for Selection Against Complex Mixtures of LigandsKarin Jacobsson0Lars Frykberg11Swedish University of Agricultural Sciences, Uppsala, Sweden1Swedish University of Agricultural Sciences, Uppsala, SwedenSelection of shotgun phage-display libraries against complex mixtures of components, such as cells or sera, may result in a high number of nonspecifically binding phage. Consequently, correct interactions may be difficult to identify. To enable discrimination between faithful and nonspecific interactions, a set of eight different gene VIII-based, phage-display vectors were constructed. All vectors contain a “universal” screening tag positioned in such a way that it is only expressed when the inserted DNA encodes an open reading frame, which corrects a shift of reading frames in the vector. A Staphylococcus aureus shotgun phagedisplay library was made in a stoichiometric mixture of all vectors. After affinity-selection against IgG, one vector completely outcompeted the others. This vector contains the promoter and signal sequence from the gene encoding staphylococcal protein A and one suppressible stop codon immediately upstream of gene VIII. An increase in the frequency of clones expressing the affinity tag in all pannings correlated with selection for ligand-binding clones. This enables detection of putatively correct clones after selection of a shotgun phagedisplay library both against purified ligands and more complex materials like calf serum.https://www.future-science.com/doi/10.2144/98242rr01 |
| spellingShingle | Karin Jacobsson Lars Frykberg Gene VIII-Based, Phage-Display Vectors for Selection Against Complex Mixtures of Ligands BioTechniques |
| title | Gene VIII-Based, Phage-Display Vectors for Selection Against Complex Mixtures of Ligands |
| title_full | Gene VIII-Based, Phage-Display Vectors for Selection Against Complex Mixtures of Ligands |
| title_fullStr | Gene VIII-Based, Phage-Display Vectors for Selection Against Complex Mixtures of Ligands |
| title_full_unstemmed | Gene VIII-Based, Phage-Display Vectors for Selection Against Complex Mixtures of Ligands |
| title_short | Gene VIII-Based, Phage-Display Vectors for Selection Against Complex Mixtures of Ligands |
| title_sort | gene viii based phage display vectors for selection against complex mixtures of ligands |
| url | https://www.future-science.com/doi/10.2144/98242rr01 |
| work_keys_str_mv | AT karinjacobsson geneviiibasedphagedisplayvectorsforselectionagainstcomplexmixturesofligands AT larsfrykberg geneviiibasedphagedisplayvectorsforselectionagainstcomplexmixturesofligands |