Document Detail


Establishment of Agrobacterium tumefaciens-mediated transformation of an oleaginous fungus, Mortierella alpina 1S-4, and its application for eicosapentaenoic acid producer breeding.
MedLine Citation:
PMID:  19581481     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Akinori Ando; Yosuke Sumida; Hiroaki Negoro; Dian Anggraini Suroto; Jun Ogawa; Eiji Sakuradani; Sakayu Shimizu
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-07-06
Journal Detail:
Title:  Applied and environmental microbiology     Volume:  75     ISSN:  1098-5336     ISO Abbreviation:  Appl. Environ. Microbiol.     Publication Date:  2009 Sep 
Date Detail:
Created Date:  2009-08-28     Completed Date:  2009-10-22     Revised Date:  2010-09-27    
Medline Journal Info:
Nlm Unique ID:  7605801     Medline TA:  Appl Environ Microbiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  5529-35     Citation Subset:  IM    
Affiliation:
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan.
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MeSH Terms
Descriptor/Qualifier:
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:
0/Fatty Acids; 1553-41-9/Eicosapentaenoic Acid; EC 1.14.19.-/Fatty Acid Desaturases; EC 1.14.99.-/omega-3 fatty acid desaturase
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