ArticleMolecules (Basel, Switzerland)2015
Application of the Cre/loxP Site-Specific Recombination System for Gene Transformation in Aurantiochytrium limacinum.
Article in Molecules (Basel, Switzerland), 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed, 12 citations in OpenAlex.
- Novel centromeric plasmid for stable extrachromosomal gene expression in Aurantiochytrium limacinum.Applied microbiology and biotechnology · 2025Article
- Deciphering and engineering the polyunsaturated fatty acid synthase pathway from eukaryotic microorganisms.Frontiers in bioengineering and biotechnology · 2022Review
- Method Development Progress in Genetic Engineering of Thraustochytrids.Marine drugs · 2021Review
- Combined Metabolome and Lipidome Analyses for In-Depth Characterization of Lipid Accumulation in the DHA ProducingMetabolites · 2021Article
- Metabolic Analysis ofFrontiers in bioengineering and biotechnology · 2021Article
- Alleviation of reactive oxygen species enhances PUFA accumulation inMetabolic engineering communications · 2018Article
- Article
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Authors and funding
9 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The Cre/loxP site-specific recombination system was applied to Aurantiochytrium limacinum to obtain a transformant without the antibiotic resistance marker gene. First, the enhanced green fluorescent protein gene (egfp) and chloramphenicol resistance gene (Cmr), along with the two loxP loci, were integrated into the genome of A. limacinum OUC88 using 18S rDNA sequences as the homologous recombination sites. Then plasmid pSH65, containing a zeocin resistance gene (Bler) was transferred into A. limacinum OUC_CG. After induction with galactose, repeated passage in culture and PCR-based assessment, the pSH65 plasmid was lost and A. limacinum OUC_EG host was shown to no longer have resistance to 100 mg chloramphenicol/L or 5 mg zeocin/L. Through southern blotting and fluorescence detection, egfp was found to be integrated into the genome of A. limacinum OUC_EG, and EGFP was successfully expressed in the cells. The successful application of the Cre/loxP system demonstrates an experimental basis for genetic modification of A. limacinum so as to obtain transformed strains with no antibiotic resistance marker genes.
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