トラマメ(Phaseolus vulgaris L.)発芽子葉α-アミラーゼcDNAのクローニング Molecular Cloning of an α-Amylase cDNA from Germinating Cotyledons of Kidney Bean (Phaseolus vulgaris L. cv. Toramame)

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Author(s)

    • 森 春英 MORI Haruhide
    • 北海道大学農学部応用生命科学科 Department of Applied Bioscience, Faculty of Agriculture, Hokkaido University
    • 松井 博和 MATSUI Hirokazu
    • 北海道大学農学部生物機能化学科 Department of Bioscience and Chemistry, Faculty of Agriculture, Hokkaido University
    • 木村 淳夫 KIMURA Atsuo
    • 北海道大学農学部応用生命科学科 Department of Applied Bioscience, Faculty of Agriculture, Hokkaido University
    • 千葉 誠哉 CHIBA Seiya
    • 北海道大学農学部応用生命科学科 Department of Applied Bioscience, Faculty of Agriculture, Hokkaido University

Abstract

トラマメ(Phaseolus vulgaris L. cv. Toramame)発芽種子の子葉由来のα一アミラーゼcDNAをクローニングし塩基配列を解析した.精製酵素の部分アミノ酸配列をもとに作成したプライマーによりPCRを行い,増幅断片をプローブとした.cDNAライブラリーは発芽種子の子葉より調製した.単離したcDNAは,1263bpのオープンリーディングフレーム(ORF),それぞれ6bpおよび194 bpの5ノおよび3ノ非翻訳領域をもつ.推定されるアミノ酸配列は,420残基からなり,N末端に23残基のシグナル配列を含む.他起源α一アミラーゼとの相同性の比較の結果,双子葉植物由来のものに70%以上,特にVigna radiate起源のものに高く94%の相同性を示した.α一アミラーゼに保存されている四つの相同性の高い領域が確認された.

A cDNA coding a-amylase of Phaseolus vulgaris L. cv. Toramame was isolated and sequenced. PCR primers were synthesized based on the partial amino acid sequences of the purified a-amylase. The DNA fragment amplified by PCR with the primers was applied as a probe to screen the cDNA library representing mRNA in the cotyledons of germinating seeds. The cDNA sequence in the positive clone encoded a unique 1263-nucleotide open reading frame flanked by 5'- and 3'-untranslated regions of 6 and 194 nucleotides, respectively. The open reading frame encoded a protein of 420 amino acid residues including a 23 amino-acid-long signal peptide. The kidney bean a-amylase shows over 70% similarity in the amino acid sequence with other dicotyledonous plant a-amylases. Especially, 94% of the residues are identical to a-amylase from Vigna radiate. Kidney bean a-amylase also has all four sequence regions highly conserved in enzymes belonging to the amylase family.

Journal

  • Journal of Applied Glycoscience

    Journal of Applied Glycoscience 45(3), 261-267, 1998-08-31

    The Japanese Society of Applied Glycoscience

References:  21

Cited by:  1

Keywords

Codes

  • NII Article ID (NAID)
    10008258281
  • NII NACSIS-CAT ID (NCID)
    AN10453916
  • Text Lang
    ENG
  • Article Type
    Journal Article
  • ISSN
    13403494
  • NDL Article ID
    4551727
  • NDL Source Classification
    ZP24(科学技術--化学・化学工業--糖・澱粉)
  • NDL Call No.
    Z17-15
  • Data Source
    CJP  CJPref  NDL  J-STAGE  JASI 
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