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Isolation and characterization of the zSSIIa and zSSIIb starch synthase cDNA clones from maize endosperm

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Abstract

Two starch synthase clones, zSSIIa and zSSIIb, were isolated from a cDNA library constructed from W64A maize endosperm. zSSIIa and zSSIIb are 3124 and 2480 bp in length, and contain open reading frames of 732 and 698 amino acid residues, respectively. The deduced amino acid sequences of the two clones share 58.1% sequence identity. Amino acid sequence identity between the zSSIIa and zSSIIb clones and the starch synthase II clones of potato and pea ranges between 45 to 51%. The predicted amino acid sequence from each SSII cDNA contains the KXGGL consensus motif at the putative ADP-Glc binding site. Both clones also contain putative transit peptides followed by the VRAA(E)A motif, the consensus cleavage site located at the C-terminus of chloroplast transit peptides. The identity of the zSSIIa and zSSIIb clones as starch synthases was confirmed by expression of enzyme activity in Escherichia coli. Genomic DNA blot analysis revealed two copies of zSSIIa and a single copy of zSSIIb. zSSIIa was expressed predominantly in the endosperm, while transcripts for zSSIIb were detected mainly in the leaf at low abundance. These findings establish that the zSSIIa and zSSIIb genes are characteristically distinct from genes encoding granule-bound starch synthase I (Waxy protein) and starch synthase I.

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Authors and Affiliations

  1. Department of Food Science, New Jersey Agricultural Experiment Station, Cook College, Rutgers University, New Brunswick, NJ, 08901-8520, USA

    Chee Harn, Aravind Ramakrishnan, Peter L. Keeling & Bruce P. Wasserman

  2. Zeneca Seeds, Berkshire, RG12 6EY, UK

    Mary Knight

  3. ExSeed Genetics, Iowa State University, Ames, IA, 50011, USA

    Hanping Guan

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  1. Chee Harn
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  2. Mary Knight
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  3. Aravind Ramakrishnan
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  4. Hanping Guan
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  5. Peter L. Keeling
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  6. Bruce P. Wasserman
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Harn, C., Knight, M., Ramakrishnan, A. et al. Isolation and characterization of the zSSIIa and zSSIIb starch synthase cDNA clones from maize endosperm. Plant Mol Biol 37, 639–649 (1998). https://doi.org/10.1023/A:1006079009072

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