1. Gene targeting in the oil-producing fungus Mortierella alpina 1S-4 and construction of a strain producing a valuable polyunsaturated fatty acid.
- Author
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Kikukawa H, Sakuradani E, Nakatani M, Ando A, Okuda T, Sakamoto T, Ochiai M, Shimizu S, and Ogawa J
- Subjects
- Arachidonic Acid metabolism, DNA metabolism, DNA Breaks, Double-Stranded, DNA End-Joining Repair, DNA, Fungal metabolism, Delta-5 Fatty Acid Desaturase, Fatty Acid Desaturases deficiency, Fatty Acid Desaturases genetics, Fungal Proteins genetics, Fungal Proteins metabolism, Genetic Engineering, Homologous Recombination, Mesylates pharmacology, Mortierella classification, Mortierella drug effects, Mortierella metabolism, Phylogeny, 8,11,14-Eicosatrienoic Acid metabolism, DNA genetics, DNA, Fungal genetics, Gene Targeting, Mortierella genetics
- Abstract
To develop an efficient gene-targeting system in Mortierella alpina 1S-4, we identified the ku80 gene encoding the Ku80 protein, which is involved in the nonhomologous end-joining pathway in genomic double-strand break (DSB) repair, and constructed ku80 gene-disrupted strains via single-crossover homologous recombination. The Δku80 strain from M. alpina 1S-4 showed no negative effects on vegetative growth, formation of spores, and fatty acid productivity, and exhibited high sensitivity to methyl methanesulfonate, which causes DSBs. Dihomo-γ-linolenic acid (DGLA)-producing strains were constructed by disruption of the Δ5-desaturase gene, encoding a key enzyme of bioconversion of DGLA to ARA, using the Δku80 strain as a host strain. The significant improvement of gene-targeting efficiency was not observed by disruption of the ku80 gene, but the construction of DGLA-producing strain by disruption of the Δ5-desaturase gene was succeeded using the Δku80 strain as a host strain. This report describes the first study on the identification and disruption of the ku80 gene in zygomycetes and construction of a DGLA-producing transformant using a gene-targeting system in M. alpina 1S-4.
- Published
- 2015
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