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Electrochemical detection of protein-protein interactions using a yeast two hybrid: 17-beta-estradiol as a model
Authors:Schwartz-Mittelmann A  Neufeld T  Biran D  Rishpon J
Institution:Department of Molecular Microbiology and Biotechnology, Tel-Aviv University, Ramat-Aviv 69978, Israel.
Abstract:In this work we present a modified yeast two-hybrid bioassay for the highly sensitive detection of protein-protein interactions, based on the electrochemical monitoring of beta-D-galactosidase reporter gene activity, using p-aminophenyl-beta-D-galactopyranoside (PAPG) as a synthetic substrate. In a model system, the sensitive detection of 17-beta-estradiol was achieved at concentrations as low as 10(-11)M (approx 2 pg/ml) by monitoring 17-beta-estradiol receptor dimerization after exposure to 17-beta-estradiol. The sensitivity of this system was higher than that of standard optical methods by three orders of magnitude.
Keywords:Yeast two hybrid  Estradiol  Electrochemical  Protein-protein interaction
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