ArticleApplied and environmental microbiology2012
Versatile transformation system that is applicable to both multiple transgene expression and gene targeting for Thraustochytrids.
Article in Applied and environmental microbiology, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed, 66 citations in OpenAlex.
- Lipid Droplet Degradation Through Lipophagy in Aurantiochytrium limacinum mh0186.Marine biotechnology (New York, N.Y.) · 2025Article
- Novel centromeric plasmid for stable extrachromosomal gene expression in Aurantiochytrium limacinum.Applied microbiology and biotechnology · 2025Article
- Enhancing fatty acid and omega-3 production in Schizochytrium sp. using developed safe-harboring expression system.Journal of biological engineering · 2024Article
- Characterization of thraustochytrid-specific sterolApplied and environmental microbiology · 2023Article
- Enhancement of Squalene Production by Constitutive Expression of the 3-Hydroxy-3-Methylglutaryl-CoA Reductase in Aurantiochytrium sp. 18W-13a.Marine biotechnology (New York, N.Y.) · 2022Article
- Deciphering and engineering the polyunsaturated fatty acid synthase pathway from eukaryotic microorganisms.Frontiers in bioengineering and biotechnology · 2022Review
- Strategic Development ofFrontiers in nutrition · 2022Article
- PUFA synthase-independent DHA synthesis pathway in Parietichytrium sp. and its modification to produce EPA and n-3DPA.Communications biology · 2021Article
- Method Development Progress in Genetic Engineering of Thraustochytrids.Marine drugs · 2021Review
- A non-canonical Δ9-desaturase synthesizing palmitoleic acid identified in the thraustochytrid Aurantiochytrium sp. T66.Applied microbiology and biotechnology · 2021Article
- Combined Metabolome and Lipidome Analyses for In-Depth Characterization of Lipid Accumulation in the DHA ProducingMetabolites · 2021Article
- Development of an Efficient Gene Editing Tool inFrontiers in nutrition · 2021Article
- Genetic tool development in marine protists: emerging model organisms for experimental cell biology.Nature methods · 2020Article
- Genome and Transcriptome Analyses Provide Insight Into the Omega-3 Long-Chain Polyunsaturated Fatty Acids Biosynthesis ofFrontiers in microbiology · 2020Article
- Enhancement of docosahexaenoic acid production by overexpression of ATP-citrate lyase and acetyl-CoA carboxylase inBiotechnology for biofuels · 2020Article
- A thraustochytrid-specific lipase/phospholipase with unique positional specificity contributes to microbial competition and fatty acid acquisition from the environment.Scientific reports · 2019Article
- Article
- Functions of PKS Genes in Lipid Synthesis of Schizochytrium sp. by Gene Disruption and Metabolomics Analysis.Marine biotechnology (New York, N.Y.) · 2018Article
- Alleviation of reactive oxygen species enhances PUFA accumulation inMetabolic engineering communications · 2018Article
- Visualization of Endoplasmic Reticulum and Mitochondria in Aurantiochytrium limacinum by the Expression of EGFP with Cell Organelle-Specific Targeting/Retaining Signals.Marine biotechnology (New York, N.Y.) · 2018Article
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Authors and funding
15 authors at 4 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A versatile transformation system for thraustochytrids, a promising producer for polyunsaturated fatty acids and fatty acid-derived fuels, was established. G418, hygromycin B, blasticidin, and zeocin inhibited the growth of thraustochytrids, indicating that multiple selectable marker genes could be used in the transformation system. A neomycin resistance gene (neo(r)), driven with an ubiquitin or an EF-1α promoter-terminator from Thraustochytrium aureum ATCC 34304, was introduced into representatives of two thraustochytrid genera, Aurantiochytrium and Thraustochytrium. The neo(r) marker was integrated into the chromosomal DNA by random recombination and then functionally translated into neo(r) mRNA. Additionally, we confirmed that another two genera, Parietichytrium and Schizochytrium, could be transformed by the same method. By this method, the enhanced green fluorescent protein was functionally expressed in thraustochytrids. Meanwhile, T. aureum ATCC 34304 could be transformed by two 18S ribosomal DNA-targeting vectors, designed to cause single- or double-crossover homologous recombination. Finally, the fatty acid Δ5 desaturase gene was disrupted by double-crossover homologous recombination in T. aureum ATCC 34304, resulting in an increase of dihomo-γ-linolenic acid (C(20:3n-6)) and eicosatetraenoic acid (C(20:4n-3)), substrates for Δ5 desaturase, and a decrease of arachidonic acid (C(20:4n-6)) and eicosapentaenoic acid (C(20:5n-3)), products for the enzyme. These results clearly indicate that a versatile transformation system which could be applicable to both multiple transgene expression and gene targeting was established for thraustochytrids.
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