Titlebook: Recombinant Protein Protocols; Detection and Isolat Rocky S. Tuan Book 1997 Springer Science+Business Media New York 1997

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Detection and Isolation of Recombinant Proteins Based on Binding Affinity of Reporter: Protein Ae first described system allowing affinity purification of expressed gene products. To date, a multitude of proteins have been produced as fusions to the IgG-binding domains of staphylococcal protein A, in several different hosts such as gram-positive and gram-negative bacteria (.,.), yeast (.), CHO cells (.), insect cells (.) and plants (.).

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978-1-4899-4167-1Springer Science+Business Media New York 1997

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Recombinant Protein Protocols978-1-59259-549-5Series ISSN 1064-3745 Series E-ISSN 1940-6029

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1064-3745 s of recombinant gene products cover a wide spectrum, including gene therapy, production of bioactive pharmaceuticals, synthesis of novel biopolymers, agriculture and animal husbandry, and so on. Inherent in bringing these appli- cations to fruition is the need to design "expression constructs" that

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Expression and Purification of Recombinant Streptavidin-Containing Chimeric Proteinsr a variety of biological and biomedical analyses (.,.). The ability of biotin to be incorporated easily into various biological materials has also expanded the application of the streptavidin-biotin technology to a wider range of biological systems.

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Detection and Isolation of Recombinant Proteins from Mammalian Cells by Immunoaffinity Chromatographpitope on the recombinant protein is mapped to a short primary sequence a synthetic epitope peptide can be used to elute the protein from the antibody under mild conditions. This technique has been employed to elute proteins from immunoaffinity columns with the retention of biological function.

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Alternative Yeast Two-Hybrid Systemsng cDNA libraries for novel interacting proteins, and screening novel proteins against predetermined sets of known proteins to identify which pairs interact. We here describe strategies for these purposes utilizing the interaction trap (.), a two-hybrid system variant developed in the laboratory of Roger Brent (.).

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Detection of Heterologous Gs-Coupled Receptor Activity in LLC-PK1 Cells Based on Expression of Urokie visualized . by overlaying colonies of stimulated cells with agar containing both plasminogen and casein (.). The degradation of the milk-protein casein by plasmin leads to the formation of lytic zones that allow the identification of uPA secreting colonies.