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Cloning and sequencing of a cDNA encoding ascorbate peroxidase fromArabidopsis thaliana

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Abstract

A cDNA clone encoding ascorbate peroxidase (AP, EC 1.11.1.11) was isolated from a phage λgt11 library of cDNA fromArabidopsis thaliana by immunoscreening with monoclonal antibodies against the enzyme, and then sequenced. The cDNA insert hybridized to a 1.1 kb poly(A)+ RNA from leaves ofA thaliana. Genomic hybridization suggests that the cDNA obtained here corresponds to a single-copy gene. The N-terminal amino acid sequence ofArabidopsis AP was determined by protein sequencing of the immunochemically purified enzyme, and proved to be homologous to the N-terminal amino acid sequence of the chloroplastic AP of spinach. The predicted amino acid sequence of the mature AP ofA. thaliana, deduced from the nucleotide sequence, consists of 249 amino acid residues, which is 34% homologous with cytochromec peroxidase of yeast, but less homologous with other plant peroxidases. Amino acid residues at the active site of yeast cytochromec peroxidase are conserved in the amino acid sequence ofArabidopsis AP. The poly(dG-dT) sequence, which is a potential Z-DNA-forming sequence, was found in the 3′ untranslated region of the cDNA.

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Authors and Affiliations

  1. Environmental Biology Division, National Institute for Environmental Studies, Onogawa, 305, Tsukuba, Ibaraki, Japan

    Akihiro Kubo & Kiyoshi Tanaka

  2. Regional and Community Environment Division, National Institute for Environmental Studies, Onogawa, 305, Tsukuba, Ibaraki, Japan

    Hikaru Saji & Noriaki Kondo

  3. Faculty of Agriculture, Kyoto Prefectural University, 606, Shimogamo, Kyoto, Japan

    Kunisuke Tanaka

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  1. Akihiro Kubo
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  2. Hikaru Saji
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  4. Kunisuke Tanaka
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  5. Noriaki Kondo
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Kubo, A., Saji, H., Tanaka, K. et al. Cloning and sequencing of a cDNA encoding ascorbate peroxidase fromArabidopsis thaliana . Plant Mol Biol 18, 691–701 (1992). https://doi.org/10.1007/BF00020011

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  • DOI: https://doi.org/10.1007/BF00020011

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