Two-Hybrid Systems Methods and Protocols /
The yeast two-hybrid system is one of the most widely used and productive techniques available for investigating the macromolecular interactions that affect virtually all biological processes. In Two-Hybrid Systems: Methods and Protocols, Paul N. MacDonald has assembled a collection of these powerfu...
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| Format: | Electronic eBook |
| Language: | English |
| Published: |
Totowa, NJ :
Humana Press : Imprint: Humana,
2001.
|
| Edition: | 1st ed. 2001. |
| Series: | Methods in Molecular Biology,
177 |
| Subjects: | |
| Online Access: | https://doi.org/10.1385/1592592104 |
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| 245 | 1 | 0 | |a Two-Hybrid Systems |h [electronic resource] : |b Methods and Protocols / |c edited by Paul N. MacDonald. |
| 250 | |a 1st ed. 2001. | ||
| 264 | 1 | |a Totowa, NJ : |b Humana Press : |b Imprint: Humana, |c 2001. | |
| 300 | |a XII, 336 p. |b online resource. | ||
| 336 | |a text |b txt |2 rdacontent | ||
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| 490 | 1 | |a Methods in Molecular Biology, |x 1064-3745 ; |v 177 | |
| 505 | 0 | |a General Methods -- The Two-Hybrid System -- Growth and Maintenance of Yeast -- Media Formulations for Various Two-Hybrid Systems -- Yeast Two-Hybrid Vectors and Strains -- High-Efficiency Transformation of Plasmid DNA into Yeast -- Qualitative and Quantitative Assessment of Interactions -- Strategies for Rescuing Plasmid DNA from Yeast Two-Hybrid Colonies -- False Positives -- Two-Hybrid System and False Positives -- Confirming Yeast Two-Hybrid Protein Interactions Using In Vitro Glutathione-S-Transferase Pulldowns -- Two-Hybrid Interactions Confirmed by Coimmunoprecipitation of Epitope-Tagged Clones -- Applications -- Smad Interactors in Bone Morphogenetic Protein Signaling -- Protein Interactions Important in Eukaryotic Translation Initiation -- Steroid Receptor and Ligand-Dependent Interaction with Coactivator Proteins -- Interaction of Cellular Apoptosis Regulating Proteins with Adenovirus Anti-apoptosis Protein E1B-19K -- Alternative Strategies -- Mammalian Two-Hybrid Assays -- One-Hybrid Systems f hor Detecting Protein-DNA Interactions -- The Split-Hybrid System -- Three-Hybrid Screens -- Three-Hybrid Screens for RNA-Binding Proteins -- Membrane Recruitment Systems for Analysis of Protein-Protein Interactions. | |
| 520 | |a The yeast two-hybrid system is one of the most widely used and productive techniques available for investigating the macromolecular interactions that affect virtually all biological processes. In Two-Hybrid Systems: Methods and Protocols, Paul N. MacDonald has assembled a collection of these powerful molecular tools for examining and characterizing protein-protein, protein-DNA, and protein-RNA interactions. The techniques range from the most basic (introducing plasmids into yeasts, interaction assays, and recovering the plasmids from yeast) to the most advanced alternative strategies (involving one-hybrid, split two-hybrid, three-hybrid, membrane recruitment systems, and mammalian systems). Methods are also provided for dealing with the well-known problems of artifacts and false positives and for identifying the interacting partners in important biological systems, including the Smad and nuclear receptor pathways. To ensure ready reproducibility and robust results, each technique is described in step-by-step detail by researchers who employ it regularly. Comprehensive and highly practical, Two-Hybrid Systems: Methods and Protocols not only reveals how the great variety of plasmid vectors and approaches may be optimally deployed, but also quickly empowers novices to establish two-hybrid systems in their laboratories, and experienced researchers to expand their repertoire of techniques. | ||
| 650 | 0 | |a Cell biology. | |
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| 700 | 1 | |a MacDonald, Paul N. |e editor. |4 edt |4 http://id.loc.gov/vocabulary/relators/edt | |
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| 830 | 0 | |a Methods in Molecular Biology, |x 1064-3745 ; |v 177 | |
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