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Detection of PERIOD/PAS-binding proteins from rat brain nuclear extracts by affinity chromatography
1Ishida Group of Clock Gene, National Institute of Bioscience and Human Technology, AIST, MITI, Tsukuba Science City, Japan.
Abstract:
In Drosophila, period and timeless gene's products (dPER and dTIM) have been identified to act as circadian clock components in heterodimeric form. The formation of dPER-dTIM complex is based on the interaction between dPER's PAS domain and dTIM's PAS binding domain. As an initial step to understand the molecular mechanism of mammalian circadian clock, we screened His-tagged dPER/PAS binding proteins from rat brain nuclear extracts, using Ni2+-NTA affinity chromatography. As a result of screening, we identified two proteins (192 and 180 kDa), which specifically bound to His-dPER/PAS in rat brain extracts. Such proteins might be a candidate of rat timeless proteins.