ArticleMarine biotechnology (New York, N.Y.)2014
Construction of novel chloroplast expression vector and development of an efficient transformation system for the diatom Phaeodactylum tricornutum.
Article in Marine biotechnology (New York, N.Y.), 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed, 80 citations in OpenAlex.
- Review
- The marine diatom Phaeodactylum tricornutum as a versatile bioproduction chassis: Current progress, challenges, and perspectives.Plant communications · 2025Review
- In Vitro Culture, Genetic Transformation and the Production of Biopharmaceuticals in Microalgae.International journal of molecular sciences · 2025Review
- Chloroplast Functionality at the Interface of Growth, Defense, and Genetic Innovation: A Multi-Omics and Technological Perspective.Plants (Basel, Switzerland) · 2025Review
- Genetic engineering in diatoms: advances and prospects.The Plant journal : for cell and molecular biology · 2025Review
- Design and assembly of the 117-kb Phaeodactylum tricornutum chloroplast genome.Plant physiology · 2024Article
- Exchange or Eliminate: The Secrets of Algal-Bacterial Relationships.Plants (Basel, Switzerland) · 2024Review
- Current Status and Perspective on the Use of Viral-Based Vectors in Eukaryotic Microalgae.Marine drugs · 2022Review
- Harnessing the Algal Chloroplast for Heterologous Protein Production.Microorganisms · 2022Review
- Stramenopile microalgae as "green biofactories" for recombinant protein production.World journal of microbiology & biotechnology · 2021Review
- Droplet-based microfluidic screening and sorting of microalgal populations for strain engineering applications.Algal research · 2021Article
- Metabolic Engineering for Carotenoid Production Using Eukaryotic Microalgae and Prokaryotic Cyanobacteria.Advances in experimental medicine and biology · 2021Article
- Cloning ofBiology · 2020Article
- Diatom Molecular Research Comes of Age: Model Species for Studying Phytoplankton Biology and Diversity.The Plant cell · 2020Review
- Bioengineering of Microalgae: Recent Advances, Perspectives, and Regulatory Challenges for Industrial Application.Frontiers in bioengineering and biotechnology · 2020Review
- Construction of a species-specific vector for improved plastid transformation efficiency in3 Biotech · 2019Article
- Constitutive and Chloroplast Targeted Expression of Acetyl-CoA Carboxylase in Oleaginous Microalgae Elevates Fatty Acid Biosynthesis.Marine biotechnology (New York, N.Y.) · 2018Article
- Genetic and metabolic engineering in diatoms.Philosophical transactions of the Royal Society of London. Series B, Biological sciences · 2017Review
- Intracellular spectral recompositioning of light enhances algal photosynthetic efficiency.Science advances · 2017Article
- Construction of Plastid Expression Vector and Development of Genetic Transformation System for the Seaweed Pyropia yezoensis.Marine biotechnology (New York, N.Y.) · 2017Article
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Authors and funding
8 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Plastids are ideal subcellular hosts for the expression of transgenes and have been successfully used for the production of different biopolymers, therapeutic proteins and industrial enzymes. Phaeodactylum tricornutum is a widely used aquatic feed species. In this study, we focused on developing a high-efficiency plastid expression system for P. tricornutum. In the plastid transformation vector, the site selected for integration was the transcriptionally active intergenic region present between the trnI and trnA genes, located in the IR (inverted repeat) regions of the plastid genome. Initially, a CAT reporter gene (encoding chloramphenicol acetyltransferase) was integrated at this site in the plastid genome. The expression of CAT in the transformed microalgae conferred resistance to the antibiotic chloramphenicol, which enabled growth in the selection media. Overall, the plastid transformation efficiency was found to be approximately one transplastomic colony per 1,000 microalgae cells. Subsequently, a heterologous gene expression cassette for high-level expression of the target gene was created and cloned between the homologous recombination elements. A TA cloning strategy based on the designed XcmI-XcmI sites could conveniently clone the heterologous gene. An eGFP (green fluorescent protein) reporter gene was used to test the expression level in the plastid system. The relatively high-level expression of eGFP without codon optimisation in stably transformed microalgae was determined to account for 0.12 % of the total soluble protein. Thus, this study presents the first and convenient plastid gene expression system for diatoms and represents an interesting tool to study diatom plastids.
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