The Two Body Problem

Two-Hybrid Systems Studying a biological phenomenon? Which field? Well, it really doesn't matter. Whatever the area of interest, rest assured that protein-protein interactions are somehow, somewhere involved--if not directly, then as part of a structural or regulatory apparatus. Two-hybrid systems, pioneered by Fields and Song,1 are powerful systems for detecting interactions between and among macromolecules. The first systems described were for protein-protein interactions; later, the basic

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Utilizing techniques common to modern molecular biology laboratories, two-hybrid systems offer a number of advantages over other biochemical methods for detecting interactions between proteins, such as immunoprecipitation, biochemical copurification, and affinity chromatography. One significant advantage is that two-hybrid systems require neither purified proteins/antibodies nor prior knowledge of a protein's function. They offer the ultimate in flexibility: Pairs of known proteins can be tested against each other for possible interactions, groups of proteins can be tested in pairwise combinations, or entire libraries of genes (cDNAs or genomic) can be tested against either an individual protein or another library, enabling the detection of completely novel sets of interacting proteins. Two-hybrid systems are marvelously efficient in that they provide the opportunity to screen entire libraries for interactions in a single experiment and in fact, have been used to map interactions within the entire genome/proteome of bacteriophage T7 of E. coli.3 A further advantage ...

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  • Laura Defrancesco

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