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KMID : 0613820050150050779
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Journal of Life Science
2005 Volume.15 No. 5 p.779 ~ p.782
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Assay of Epoxide Hydrolase Activity Based on PCR-linked in vitro Coupled Transcription and Translation System
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Lee Ok-Kyung
Kim Hee-Sook
Lee Eun-Yeol
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Abstract
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Cell-free expression is a powerful tool for rapid protein analysis, enabling an efficient identification of gene without cumbersome procedure of transformation and cell culture. Epoxide hydrolase (EH) gene of Rhodotorula glutinis was simply amplified by PCR, and the resultant gene was expressed in vitro using a coupled transcription/translation system. The cell-free expressed EH protein mixture exhibited the enantioselective hydrolysis activity toward (R)-styrene oxide, representing that cell-free protein synthesis system can be used for the rapid expression of an enantioselective enzyme for an efficient identification of the chiral activity.
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