ArticleFrontiers in bioengineering and biotechnology2016
Refinement of the Diatom Episome Maintenance Sequence and Improvement of Conjugation-Based DNA Delivery Methods.
Article in Frontiers in bioengineering and biotechnology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 40 papers.
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40 citing papers in PubMed, 90 citations in OpenAlex.
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- Pan-microalgal dark proteome mapping via interpretable deep learning and synthetic chimeras.Patterns (New York, N.Y.) · 2025Article
- The marine diatom Phaeodactylum tricornutum as a versatile bioproduction chassis: Current progress, challenges, and perspectives.Plant communications · 2025Review
- Both major xanthophyll cycles present in nature promote nonphotochemical quenching in a model diatom.Plant physiology · 2025Article
- Novel centromeric plasmid for stable extrachromosomal gene expression in Aurantiochytrium limacinum.Applied microbiology and biotechnology · 2025Article
- Characterization of the VOC Promoter That Is Active Under Low-Salinity Conditions in the DiatomMarine drugs · 2025Article
- Genetic engineering in diatoms: advances and prospects.The Plant journal : for cell and molecular biology · 2025Review
- Metabolic response to a heterologous poly-3-hydroxybutyrate (PHB) pathway in Phaeodactylum tricornutum.Applied microbiology and biotechnology · 2024Article
- Genome-Wide Mapping of Autonomously Replicating Sequences in the Marine Diatom Phaeodactylum tricornutum.Marine biotechnology (New York, N.Y.) · 2024Article
- No two clones are alike: characterization of heterologous subpopulations in a transgenic cell line of the model diatom Phaeodactylum tricornutum.Microbial cell factories · 2024Article
- Tuning interdomain conjugation to enablemSystems · 2024Article
- Chromosome-scale assembly of the streamlined picoeukaryoteMicrobial genomics · 2024Article
- Article
- Targeted Gene Editing of Nuclear-Encoded Plastid Proteins in Phaeodactylum tricornutum via CRISPR/Cas9.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Validating a Promoter Library for Application in Plasmid-Based Diatom Genetic Engineering.ACS synthetic biology · 2023Article
- Chromosome-Scale Genome Assembly of the Marine Oleaginous Diatom Fistulifera solaris.Marine biotechnology (New York, N.Y.) · 2022Article
- Phosphate-regulated expression of the SARS-CoV-2 receptor-binding domain in the diatom Phaeodactylum tricornutum for pandemic diagnostics.Scientific reports · 2022Article
- Genome editing with removable TALEN vectors harboring a yeast centromere and autonomous replication sequence in oleaginous microalga.Scientific reports · 2022Article
Corrections and comments
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Authors and funding
5 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Conjugation of episomal plasmids from bacteria to diatoms advances diatom genetic manipulation by simplifying transgene delivery and providing a stable and consistent gene expression platform. To reach its full potential, this nascent technology requires new optimized expression vectors and a deeper understanding of episome maintenance. Here, we present the development of an additional diatom vector (pPtPBR1), based on the parent plasmid pBR322, to add a plasmid maintained at medium copy number in Escherichia coli to the diatom genetic toolkit. Using this new vector, we evaluated the contribution of individual yeast DNA elements comprising the 1.4-kb tripartite CEN6-ARSH4-HIS3 sequence that enables episome maintenance in Phaeodactylum tricornutum. While various combinations of these individual elements enable efficient conjugation and high exconjugant yield in P. tricornutum, individual elements alone do not. Conjugation of episomes containing CEN6-ARSH4 and a small sequence from the low GC content 3' end of HIS3 produced the highest number of diatom exconjugant colonies, resulting in a smaller and more efficient vector design. Our findings suggest that the CEN6 and ARSH4 sequences function differently in yeast and diatoms, and that low GC content regions of greater than ~500 bp are a potential indicator of a functional diatom episome maintenance sequence. Additionally, we have developed improvements to the conjugation protocol including a high-throughput option utilizing 12-well plates and plating methods that improve exconjugant yield and reduce time and materials required for the conjugation protocol. The data presented offer additional information regarding the mechanism by which the yeast-derived sequence enables diatom episome maintenance and demonstrate options for flexible vector design.
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