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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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Published in: | Biological & pharmaceutical bulletin 2006, Vol.29(7), pp.1314-1318 |
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creator | Kubota, Takaaki Iinuma, Yoshiro Kobayashi, Jun'ichi |
description | 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). |
doi_str_mv | 10.1248/bpb.29.1314 |
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(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).</description><subject>Amphidinium</subject><subject>Amphidinium sp</subject><subject>amphidinolide</subject><subject>Animals</subject><subject>Cloning, Molecular</subject><subject>dinoflagellate</subject><subject>Dinoflagellida - enzymology</subject><subject>DNA - genetics</subject><subject>DNA - isolation & purification</subject><subject>Macrolides - metabolism</subject><subject>polyketide synthase</subject><subject>Polyketide Synthases - genetics</subject><subject>Polyketide Synthases - metabolism</subject><subject>Polymerase Chain Reaction</subject><subject>Protein Biosynthesis</subject><subject>Recombinant Proteins - metabolism</subject><issn>0918-6158</issn><issn>1347-5215</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><recordid>eNqFkUFv1DAQhS0EotuFE3cUCYkLynbGcRL7RrXQUqlSKwFny3Em3SxJHOzksP8eR7ssEpdextL489Pze4y9Q9ggF_KqGqsNVxvMULxgK8xEmeYc85dsBQplWmAuL9hlCHsAKIFnr9kFFhIVFrBietu5oR2eEtckj647_KKprSn5fhimnQmU3NJAIWm865Prfty1dTu4LhLpo3f1bJeXX-Kq6cwTdZ2Z6Iy1c5-E8Q171Zgu0NvTuWY_b77-2H5L7x9u77bX96nNBU5pDVxRndccELKyqhtpqzwXmTIkZQOCFMgsGraccwOAghdVjsSFlVQA5tmafTzqjt79nilMum-DXSwN5OagC1lAGWN5FkQlFAIXEfzwH7h3sx_iJzQKobICRPS6Zp-OlPUuBE-NHn3bG3_QCHqpR8d6NFd6qSfS70-ac9VT_Y899RGBz0dgH6aY6BkwfmptR3_FyuNYNM9Xdme8piH7AxCXoNs</recordid><startdate>20060701</startdate><enddate>20060701</enddate><creator>Kubota, Takaaki</creator><creator>Iinuma, Yoshiro</creator><creator>Kobayashi, Jun'ichi</creator><general>The Pharmaceutical Society of Japan</general><general>Japan Science and Technology Agency</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QP</scope><scope>7QR</scope><scope>7TK</scope><scope>7U9</scope><scope>8FD</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>M7N</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20060701</creationdate><title>Cloning of Polyketide Synthase Genes from Amphidinolide-Producing Dinoflagellate Amphidinium sp</title><author>Kubota, Takaaki ; Iinuma, Yoshiro ; Kobayashi, Jun'ichi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c541t-d029ed5d201037bdf8cb55439ae88f04e9083160c222a001426b51e24c8e60153</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Amphidinium</topic><topic>Amphidinium sp</topic><topic>amphidinolide</topic><topic>Animals</topic><topic>Cloning, Molecular</topic><topic>dinoflagellate</topic><topic>Dinoflagellida - enzymology</topic><topic>DNA - genetics</topic><topic>DNA - isolation & purification</topic><topic>Macrolides - metabolism</topic><topic>polyketide synthase</topic><topic>Polyketide Synthases - genetics</topic><topic>Polyketide Synthases - metabolism</topic><topic>Polymerase Chain Reaction</topic><topic>Protein Biosynthesis</topic><topic>Recombinant Proteins - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kubota, Takaaki</creatorcontrib><creatorcontrib>Iinuma, Yoshiro</creatorcontrib><creatorcontrib>Kobayashi, Jun'ichi</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Biological & pharmaceutical bulletin</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kubota, Takaaki</au><au>Iinuma, Yoshiro</au><au>Kobayashi, Jun'ichi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cloning of Polyketide Synthase Genes from Amphidinolide-Producing Dinoflagellate Amphidinium sp</atitle><jtitle>Biological & pharmaceutical bulletin</jtitle><addtitle>Biol Pharm Bull</addtitle><date>2006-07-01</date><risdate>2006</risdate><volume>29</volume><issue>7</issue><spage>1314</spage><epage>1318</epage><pages>1314-1318</pages><issn>0918-6158</issn><eissn>1347-5215</eissn><abstract>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).</abstract><cop>Japan</cop><pub>The Pharmaceutical Society of Japan</pub><pmid>16819160</pmid><doi>10.1248/bpb.29.1314</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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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 |
title | Cloning of Polyketide Synthase Genes from Amphidinolide-Producing Dinoflagellate Amphidinium sp |
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