ArticleJournal of biological engineering2013
Assembly of eukaryotic algal chromosomes in yeast.
Article in Journal of biological engineering, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 37 papers.
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Who cites it
37 citing papers in PubMed, 65 citations in OpenAlex.
- Advances in large DNA fragment assembly for microbial cell factory engineering.Quantitative biology (Beijing, China) · 2026Review
- One Yeast, Sixteen Synthetic Chromosomes, Infinite Possibilities.Yeast (Chichester, England) · 2026Review
- Building synthetic chromosomes one yeast at a time: insights from Sc2.0.Nature biotechnology · 2025Article
- Impact of heterologous expression of Cannabis sativa tetraketide synthase on Phaeodactylum tricornutum metabolic profile.Biotechnology for biofuels and bioproducts · 2025Article
- Genetic engineering in diatoms: advances and prospects.The Plant journal : for cell and molecular biology · 2025Review
- Cloning a Chloroplast Genome inBio-protocol · 2025Article
- Review
- Bioengineering of the Marine DiatomInternational journal of molecular sciences · 2023Article
- Combining Fusion of Cells with CRISPR-Cas9 Editing for the Cloning of Large DNA Fragments or Complete Bacterial Genomes in Yeast.ACS synthetic biology · 2023Article
- Enabling technology and core theory of synthetic biology.Science China. Life sciences · 2023Review
- Cross-species microbial genome transfer: a Review.Frontiers in bioengineering and biotechnology · 2023Review
- Enzymatic Supercoiling of Bacterial Chromosomes Facilitates Genome Manipulation.ACS synthetic biology · 2022Article
- Phosphate-regulated expression of the SARS-CoV-2 receptor-binding domain in the diatom Phaeodactylum tricornutum for pandemic diagnostics.Scientific reports · 2022Article
- Biocircuits in plants and eukaryotic algae.Frontiers in plant science · 2022Review
- Telomere-to-telomere genome assembly ofPeerJ · 2022Article
- Synthetic Genomics From a Yeast Perspective.Frontiers in bioengineering and biotechnology · 2022Review
- Highly Selective, CRISPR/Cas9-Mediated Isolation of Genes and Genomic Loci from Complex Genomes by TAR Cloning in Yeast.Current protocols · 2021Article
- Droplet-based microfluidic screening and sorting of microalgal populations for strain engineering applications.Algal research · 2021Article
- An artificial chromosome for data storage.National science review · 2021Article
- A supernumerary designer chromosome for modular in vivo pathway assembly in Saccharomyces cerevisiae.Nucleic acids research · 2021Article
Corrections and comments
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Authors and funding
16 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundSynthetic genomic approaches offer unique opportunities to use powerful yeast and Escherichia coli genetic systems to assemble and modify chromosome-sized molecules before returning the modified DNA to the target host. For example, the entire 1 Mb Mycoplasma mycoides chromosome can be stably maintained and manipulated in yeast before being transplanted back into recipient cells. We have previously demonstrated that cloning in yeast of large (> ~ 150 kb), high G + C (55%) prokaryotic DNA fragments was improved by addition of yeast replication origins every ~100 kb. Conversely, low G + C DNA is stable (up to at least 1.8 Mb) without adding supplemental yeast origins. It has not been previously tested whether addition of yeast replication origins similarly improves the yeast-based cloning of large (>150 kb) eukaryotic DNA with moderate G + C content. The model diatom Phaeodactylum tricornutum has an average G + C content of 48% and a 27.4 Mb genome sequence that has been assembled into chromosome-sized scaffolds making it an ideal test case for assembly and maintenance of eukaryotic chromosomes in yeast.
resultsWe present a modified chromosome assembly technique in which eukaryotic chromosomes as large as ~500 kb can be assembled from cloned ~100 kb fragments. We used this technique to clone fragments spanning P. tricornutum chromosomes 25 and 26 and to assemble these fragments into single, chromosome-sized molecules. We found that addition of yeast replication origins improved the cloning, assembly, and maintenance of the large chromosomes in yeast. Furthermore, purification of the fragments to be assembled by electroelution greatly increased assembly efficiency.
conclusionsEntire eukaryotic chromosomes can be successfully cloned, maintained, and manipulated in yeast. These results highlight the improvement in assembly and maintenance afforded by including yeast replication origins in eukaryotic DNA with moderate G + C content (48%). They also highlight the increased efficiency of assembly that can be achieved by purifying fragments before assembly.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.