2010Journal of the Faculty of Agriculture Kyushu UniversityOpen access

Molecular Cloning of the Genes Involved in Anthocyanin Biosynthesis in Camellia japonica

Nobumine Tateishi, Yukio Ozaki, Hiroshi Okubo

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Abstract

Partial cDNA sequences of three anthocyanin biosynthetic genes (F3H, flavanone 3-hydroxylase; DFR, dihydroflavonol 4-reductase; ANS, anthocyanidin synthase) were isolated from the petals of Camellia japonica. Their deduced partial amino acid sequences shared high homologies with those of woody plant species (CjF3Ha, 98.0%, CjF3Hb, 91.2% and CjDFR, 99.0% with Camellia senensis; CjANS, 90.3% with Rhododendron × pulchrum). Some important amino acid residues for enzymatic activities were also conserved in the isolated clones, suggesting that the genes we identified in this study were the homologues of C. japonica. Gene-specific primer pairs were designed based on each partial cDNA sequence. The application of these primer pairs to RT-PCR analyses was tested.

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Partial cDNA sequences of three anthocyanin biosynthetic genes (F3H, flavanone 3-hydroxylase; DFR, dihydroflavonol 4-reductase; ANS, anthocyanidin synthase) were isolated from the petals of Camellia japonica. Their deduced partial amino acid sequences shared high homologies with those of woody plant species (CjF3Ha, 98.0%, CjF3Hb, 91.2% and CjDFR, 99.0% with Camellia senensis; CjANS, 90.3% with Rhododendron × pulchrum). Some important amino acid residues for enzymatic activities were also conserved in the isolated clones, suggesting that the genes we identified in this study were the homologues of C. japonica. Gene-specific primer pairs were designed based on each partial cDNA sequence. The application of these primer pairs to RT-PCR analyses was tested.

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Available abstract

Partial cDNA sequences of three anthocyanin biosynthetic genes (F3H, flavanone 3-hydroxylase; DFR, dihydroflavonol 4-reductase; ANS, anthocyanidin synthase) were isolated from the petals of Camellia japonica. Their deduced partial amino acid sequences shared high homologies with those of woody plant species (CjF3Ha, 98.0%, CjF3Hb, 91.2% and CjDFR, 99.0% with Camellia senensis; CjANS, 90.3% with Rhododendron × pulchrum). Some important amino acid residues for enzymatic activities were also conserved in the isolated clones, suggesting that the genes we identified in this study were the homologues of C. japonica. Gene-specific primer pairs were designed based on each partial cDNA sequence. The application of these primer pairs to RT-PCR analyses was tested.

Key concepts: Cloning (programming), Gene, Genetics, Biology, Molecular cloning, Japonica, Botany, Complementary DNA

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