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77953873053
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note
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A coding region for a full-length GA2ox2 gene (Clone Name: U20502) was purchased from ABRC, and amplified by polymerase chain reaction with specific primers to its full-length cDNA sequences and suitable restriction enzyme sites (SacI and XhoI) at both ends of each primer. After a confirmation by sequence analyses, the cDNA fragment of GA2ox2 gene was ligated to pET32a vector (Novagen/Merck Biosciences, Madison, WI, USA). The preparation of recombinant GA2ox2 was confirmed on a SDS-PAGE profile, compared with a profile from negative control fraction (vector only). According to the manufacturer's procedure, affinity purification of GA2ox2 was performed by using its His-tag (Clontech TARON metal affinity resin, Takara Bio, Shiga, Japan).
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0037256863
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34249699621
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Yamauchi Y., Takeda-Kamiya N., Hanada A., Ogawa M., Kuwahara A., Seo M., Kamiya Y., and Yamaguchi S. Plant Cell Physiol. 48 (2007) 555
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Yamaguchi, S.8
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77953872897
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note
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-1, He flow: 1.0 ml/min, ion source temperature: 280 °C, ionization: EI (70 eV)]. Each metabolite was identified by comparing its MS fragment pattern and Kovats' retention index (KRI) to those of authentic specimen.
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77953872140
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note
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3/EtOAc/AcOH (20:20:1, v/v/v). Radioactivity was visualized with an imaging plate (BAS-TR2040, Fuji Photo Film, Tokyo, Japan) and analyzer (FLA-5000, Fuji Photo Film). Quantification of metabolites and substrate that remained were estimated as pixels by scanning the imaging spots with CS Analyzer (Version 3.00.1010, ATTO, Tokyo).
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note
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-1).
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