Affiliation:
1. Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan
2. Research Division of Microbial Sciences, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan
Abstract
ABSTRACT
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 ω3-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 ω3-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.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Cited by
89 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献