ArticleProceedings of the National Academy of Sciences of the United States of America1996
Specific cloning of human DNA as yeast artificial chromosomes by transformation-associated recombination.
Article in Proceedings of the National Academy of Sciences of the United States of America, 1996. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 76 papers.
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Who cites it
76 citing papers in PubMed, 159 citations in OpenAlex.
- Advances in large DNA fragment assembly for microbial cell factory engineering.Quantitative biology (Beijing, China) · 2026Review
- Synthetic Chromosomes in Yeast: A Platform for Understanding and Programming Eukaryotic Life.Microbial biotechnology · 2026Review
- A Rapid and Cost-Effective Pipeline to Identify and Capture BGCs From Bacterial Draft Genomes.Bio-protocol · 2025Article
- Development of GFP-expressing infectious clones for PRRSV using TAR cloning for antiviral drug screening.Npj viruses · 2025Article
- Assembly of ascovirus HvAV-3h long DNA fragment using the Transformation-Associated Recombination (TAR) approach in yeast cells.BMC biotechnology · 2025Article
- Development of viral infectious clones and their applications based on yeast and bacterial artificial chromosome platforms.Molecular biomedicine · 2025Review
- Heterologous expression facilitates the discovery and characterization of marine microbial natural products.Engineering microbiology · 2024Review
- Robust and Reproducible Protocol for Phage Genome "Rebooting" Using Transformation-Associated Recombination (TAR) Cloning into Yeast Centromeric Plasmid.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Efficient assembly of a synthetic attenuated SARS-CoV-2 genome inJournal of genetics · 2024Review
- Review
- 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
- Recent advances in the direct cloning of large natural product biosynthetic gene clusters.Engineering microbiology · 2023Review
- YLC-assembly: large DNA assembly via yeast life cycle.Nucleic acids research · 2023Article
- Identification, Heterologous Expression, and Characterization of the Tolypodiol Biosynthetic Gene Cluster through an Integrated Approach.ACS chemical biology · 2023Article
- Miramides A-D: Identification of Detoxin-like Depsipeptides after Heterologous Expression of a Hybrid NRPS-PKS Gene Cluster fromMicroorganisms · 2022Article
- Multiplying the heterologous production of spinosad through tandem amplification of its biosynthetic gene cluster in Streptomyces coelicolor.Microbial biotechnology · 2022Article
- Synthetic Genomics From a Yeast Perspective.Frontiers in bioengineering and biotechnology · 2022Review
- In-yeast reconstruction of the African swine fever virus genome isolated from clinical samples.STAR protocols · 2021Article
- Saccharomyces cerevisiae as host for the recombinant production of polyketides and nonribosomal peptides.Microbial cell factories · 2021Review
- Highly Selective, CRISPR/Cas9-Mediated Isolation of Genes and Genomic Loci from Complex Genomes by TAR Cloning in Yeast.Current protocols · 2021Article
16 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
Abstract
DNA molecules undergoing transformation into yeast are highly recombinogenic, even when diverged. We reasoned that transformation-associated recombination (TAR) could be employed to clone large DNAs containing repeat sequences, thereby eliminating the need for in vitro enzymatic reactions such as restriction and ligation and reducing the amount of DNA handling. Gently isolated human DNA was transformed directly into yeast spheroplasts along with two genetically marked (M1 and M2) linearized vectors that contained a human Alu sequence at one end and a telomere sequence at the other end (Alu-CEN-M1-TEL and Alu-M2-TEL). Nearly all the M1-selected transformants had yeast artificial chromosomes (YACs) containing human DNA inserts that varied in size from 70 kb to > 600 kb. Approximately half of these had also acquired the unselected M2 marker. The mitotic segregational stability of YACs generated from one (M1) or two (M1 and M2) vector(s) was comparable, suggesting de novo generation of telomeric ends. Since no YACs were isolated when rodent DNAs or a vector lacking an Alu sequence was used, the YACs were most likely the consequence of TAR between the repeat elements on the vector(s) and the human DNA. Using the BLUR13 Alu-containing vector, we demonstrated that human DNA could be efficiently cloned from mouse cells that contained a single human chromosome 16. The distribution of cloned DNAs on chromosome 16 was determined by fluorescence in situ hybridization. We propose that TAR cloning can provide an efficient means for generating YACs from specific chromosomes and subchromosome fragments and that TAR cloning may be useful for isolating families of genes and specific genes from total genome DNA.
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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.