ArticleNucleic acids research2003
Optimum conditions for selective isolation of genes from complex genomes by transformation-associated recombination cloning.
Article in Nucleic acids research, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed.
- A simple method for rapid cloning of complete herpesvirus genomes.Cell reports methods · 2024Article
- 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
- Highly Selective, CRISPR/Cas9-Mediated Isolation of Genes and Genomic Loci from Complex Genomes by TAR Cloning in Yeast.Current protocols · 2021Article
- Metathramycin, a new bioactive aureolic acid discovered by heterologous expression of a metagenome derived biosynthetic pathway.RSC chemical biology · 2021Article
- The genomic structure of a human chromosome 22 nucleolar organizer region determined by TAR cloning.Scientific reports · 2021Article
- A Family of Single Copy repABC-Type Shuttle Vectors Stably Maintained in the Alpha-Proteobacterium Sinorhizobium meliloti.ACS synthetic biology · 2017Article
- One-step assembly and targeted integration of multigene constructs assisted by the I-SceI meganuclease in Saccharomyces cerevisiae.FEMS yeast research · 2013Article
- Parallel in vivo DNA assembly by recombination: experimental demonstration and theoretical approaches.PloS one · 2013Article
- Cloning whole bacterial genomes in yeast.Nucleic acids research · 2010Article
- Synthesis of DNA fragments in yeast by one-step assembly of overlapping oligonucleotides.Nucleic acids research · 2009Article
- Selective isolation of genomic loci from complex genomes by transformation-associated recombination cloning in the yeast Saccharomyces cerevisiae.Nature protocols · 2008Article
- Dynamic structure of the SPANX gene cluster mapped to the prostate cancer susceptibility locus HPCX at Xq27.Genome research · 2005Article
- Rapid generation of long synthetic tandem repeats and its application for analysis in human artificial chromosome formation.Nucleic acids research · 2005Article
- Closing the gaps on human chromosome 19 revealed genes with a high density of repetitive tandemly arrayed elements.Genome research · 2004Article
- A general cloning system to selectively isolate any eukaryotic or prokaryotic genomic region in yeast.BMC genomics · 2003Article
- Separation of long-range human TERT gene haplotypes by transformation-associated recombination cloning in yeast.Oncogene · 2003Article
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6 authors.
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No grant is acknowledged in the PubMed record.
Abstract
Transformation-associated recombination (TAR) cloning in yeast is used to isolate a desired chromosomal region or gene from a complex genome without construction of a genomic library. The technique involves homologous recombination during yeast spheroplast transformation between genomic DNA and a TAR vector containing short 5' and 3' gene-specific targeting hooks. Efficient gene capture requires a high yield of transformants, and we demonstrate here that the transformant yield increases approximately 10-fold when the genomic DNA is sheared to 100-200 kb before being presented to the spheroplasts. Here we determine the most effective concentration of genomic DNA, and also show that the targeted sequences recombine much more efficiently with the vector's targeting hooks when they are located at the ends of the genomic DNA fragment. We demonstrate that the yield of gene-positive clones increases approximately 20-fold after endonuclease digestion of genomic DNA, which caused double strand breaks near the targeted sequences. These findings have led to a greatly improved protocol.
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