| Establishment of Agrobacterium tumefaciens-mediated transformation of an oleaginous fungus, Mortierella alpina 1S-4, and its application for eicosapentaenoic acid producer breeding. | |
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MedLine Citation:
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PMID: 19581481 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Gene manipulation tools for an arachidonic-producing filamentous fungus, Mortierella alpina 1S-4, have not been sufficiently developed. In this study, Agrobacterium tumefaciens-mediated transformation (ATMT) was investigated for M. alpina 1S-4 transformation, using the uracil-auxotrophic mutant (ura5(-) strain) of M. alpina 1S-4 as a host strain and the homologous ura5 gene as a selectable marker gene. Furthermore, the gene for omega3-desaturase, catalyzing the conversion of n-6 fatty acid to n-3 fatty acid, was overexpressed in M. alpina 1S-4 by employing the ATMT system. As a result, we revealed that the frequency of transformation surpassed 400 transformants/10(8) spores, most of the integrated T-DNA appeared as a single copy at a random position in chromosomal DNA, and most of the transformants (60 to 80%) showed mitotic stability. Moreover, the accumulation of n-3 fatty acid in transformants was observed under the conditions of optimal omega3-desaturase gene expression. In particular, eicosapentaenoic acid (20:5n-3), an end product of n-3 fatty acids synthesized in M. alpina 1S-4, reached a maximum of 40% of total fatty acids. In conclusion, the ATMT system was found to be effective and suitable for the industrial strain Mortierella alpina 1S-4 and will be a useful tool for basic mutagenesis research and for industrial breeding of this strain. |
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Authors:
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Akinori Ando; Yosuke Sumida; Hiroaki Negoro; Dian Anggraini Suroto; Jun Ogawa; Eiji Sakuradani; Sakayu Shimizu |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2009-07-06 |
Journal Detail:
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Title: Applied and environmental microbiology Volume: 75 ISSN: 1098-5336 ISO Abbreviation: Appl. Environ. Microbiol. Publication Date: 2009 Sep |
Date Detail:
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Created Date: 2009-08-28 Completed Date: 2009-10-22 Revised Date: 2010-09-27 |
Medline Journal Info:
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Nlm Unique ID: 7605801 Medline TA: Appl Environ Microbiol Country: United States |
Other Details:
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Languages: eng Pagination: 5529-35 Citation Subset: IM |
Affiliation:
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Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Chromosomes, Fungal Eicosapentaenoic Acid / biosynthesis* Fatty Acid Desaturases / genetics Fatty Acids / metabolism Gene Dosage Gene Transfer Techniques* Genetic Engineering / methods* Genomic Instability Mortierella / genetics* Recombination, Genetic Rhizobium radiobacter / genetics* Transformation, Genetic* |
| Chemical | |
Reg. No./Substance:
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0/Fatty Acids; 1553-41-9/Eicosapentaenoic Acid; EC 1.14.19.-/Fatty Acid Desaturases; EC 1.14.99.-/omega-3 fatty acid desaturase |
| Comments/Corrections | |
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