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생화학분자생물학회 (Korean Society for Biochemistry and Molecular Biology)
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Molecular Cloning and Characterization of a New cDNA Encoding Hyoscyamine 6β-hydroxylase from Roots of Anisodus acutangulus

  • Kai, Guoyin (Laboratory of Plant Biotechnology, College of Life and Environment Sciences, Shanghai Normal University) ;
  • Chen, Junfeng (Laboratory of Plant Biotechnology, College of Life and Environment Sciences, Shanghai Normal University) ;
  • Li, Li (Laboratory of Plant Biotechnology, College of Life and Environment Sciences, Shanghai Normal University) ;
  • Zhou, Genyu (Laboratory of Plant Biotechnology, College of Life and Environment Sciences, Shanghai Normal University) ;
  • Zhou, Limin (Laboratory of Plant Biotechnology, College of Life and Environment Sciences, Shanghai Normal University) ;
  • Zhang, Lei (School of Pharmacy, Second Military Medical University) ;
  • Chen, Yuhui (Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiaotong University) ;
  • Zhao, Linxia (Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiaotong University)
  • 발행 : 2007.09.30
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초록

A new full-length cDNA encoding hyoscyamine $6\beta$-hydroxylase (designated as aah6h, GenBank Accession No. EF187826), which catalyzes the last committed step in the scopolamine biosynthetic pathway, was isolated from young roots of Anisodus acutangulus by rapid amplification of cDNA ends (RACE) for the first time. The full-length cDNA of aah6h was 1380 bp and contained a 1035 bp open reading frame (ORF) encoding a deduced protein of 344 amino acid residues. The deduced protein had an isoelectric point (pI) of 5.09 and a calculated molecular mass of about 38.7 kDa. Sequence analyses showed that AaH6H had high homology with other H6Hs isolated from some scopolamine-producing plants such as Hyoscyamus niger, Datura metel and Atropa belladonna etc. Bioinformatics analyses results indicated AaH6H belongs to 2-oxoglutarate-dependent dioxygenase superfamily. Phylogenetic tree analysis showed that AaH6H had closest relationship with H6H from A. tanguticus. Southern hybridization analysis of the genomic DNA revealed that aah6h belonged to a multi-copy gene family. Tissue expression pattern analysis firstly founded that aah6h expressed in all the tested tissues including roots, stems and leaves and indicated that aah6h was a constitutive-expression gene, which was the first reported tissue-independent h6h gene compared to other known h6h genes.

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참고문헌

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