Cloning of Polyketide Synthase Genes from Amphidinolide-Producing Dinoflagellate Amphidinium sp

Cloning of polyketide synthase (PKS) gene for amphidinolide biosynthesis was attempted from a dinoflagellate Amphidinium sp. (strain Y-42). Fourteen β-ketoacyl synthase gene fragments were obtained by Polymerase Chain Reaction (PCR) amplification from degenerated primer sets designed on the basis of...

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Bibliographic Details
Published in:Biological & pharmaceutical bulletin 2006, Vol.29(7), pp.1314-1318
Main Authors: Kubota, Takaaki, Iinuma, Yoshiro, Kobayashi, Jun'ichi
Format: Article
Language:English
Subjects:
Amphidinium
Amphidinium sp
amphidinolide
Animals
Cloning, Molecular
dinoflagellate
Dinoflagellida - enzymology
DNA - genetics
DNA - isolation & purification
Macrolides - metabolism
polyketide synthase
Polyketide Synthases - genetics
Polyketide Synthases - metabolism
Polymerase Chain Reaction
Protein Biosynthesis
Recombinant Proteins - metabolism
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Summary:Cloning of polyketide synthase (PKS) gene for amphidinolide biosynthesis was attempted from a dinoflagellate Amphidinium sp. (strain Y-42). Fourteen β-ketoacyl synthase gene fragments were obtained by Polymerase Chain Reaction (PCR) amplification from degenerated primer sets designed on the basis of the conserved amino acid sequences of β-ketoacyl synthase domains in known type I PKSs. The PCR analysis using primer sets designed from these fourteen β-ketoacyl synthase gene fragments revealed that these DNA sequences exist only in the dinoflagellates producing amphidinolides. The DNA sequence of the positive clone, which was isolated from genomic DNA library of Amphidinium sp. (strain Y-42) by PCR detection using the specific primer set, was analyzed by shotgun sequencing. The deduced gene products in the positive clone showed similarity with β-ketoacyl synthase (KS), acyl transferase (AT), dehydratase (DH), ketoreductase (KR), and acyl carrier protein (ACP) in known type I PKSs and thioesterase (TE).
ISSN:0918-6158
1347-5215
DOI:10.1248/bpb.29.1314