Abstract
The structural and functional analysis of mammalian genomes would benefit from the ability to isolate from multiple DNA samples any targeted chromosomal segment that is the size of an average human gene. A cloning technique that is based on transformation-associated recombination (TAR) in the yeast Saccharomyces cerevisiae satisfies this need. It is a unique tool to selectively recover chromosome segments that are up to 250 kb in length from complex genomes. In addition, TAR cloning can be used to characterize gene function and genome variation, including polymorphic structural rearrangements, mutations and the evolution of gene families, and for long-range haplotyping.
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Glossary
- Alphoid DNA
-
A 170 bp AT-rich repeat that is present in the centromeric regions of all human chromosomes. The repeat forms large tandem arrays with sizes of up to 5 Mb.
- Alu interspersed
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A ∼300 bp repeat that is specific to primates. There are approximately 1.5 million copies of this repeat in the human genome.
- ARS
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Autonomously replicating sequence — a small DNA fragment that can support replication in the budding yeast.
- Comparative genomic hybridization
-
A technology through which tumour and reference DNA are differentially labelled to show copy-number changes in tumour genomes.
- Contig
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From contiguous. A set of clones that represent a continuous region of DNA. A contig map depicts the relative order of a linked library of contigs that represent a complete chromosome segment.
- HAC
-
Human artificial chromosome — a system for cloning large DNA segments in mammalian cells. Similar to natural chromosomes, HACs have a functional centromere and therefore are stably maintained in a single copy during mitotic cell divisions.
- Kinetochore
-
The protein structure in eukaryotes that assembles on the centromere and links the chromosome to microtubule polymers from the mitotic spindle during mitosis.
- Minisatellites
-
A class of repetitive sequences, each of which is 7–100 nucleotides, that span 500–20,000 bp and are mainly located towards chromosome ends.
- Segmental duplications
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Large genomic segments of recent origin and nearly identical sequence.
- Spermatocyte
-
A male gametocyte that is derived from a spermatogonium. Initially, a spermatogonium divides by mitosis into two 'primary spermatocytes'. A primary spermatocyte is diploid and, by meiosis, gives place to two secondary spermatocytes, which are haploid.
- Synteny
-
Collinearity in the order of genes (or of other DNA sequences) in a chromosomal region of two species.
- Yeast spheroplasts
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Yeast cells following the removal of the cell wall. They can take up large DNA molecules (up to 1 Mb), and were used to construct the first genomic YAC libraries.
- Z-DNA
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A form of DNA in which the double helix winds to the left in a zig-zag pattern (instead of to the right, like the more common B-DNA form).
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Kouprina, N., Larionov, V. TAR cloning: insights into gene function, long-range haplotypes and genome structure and evolution. Nat Rev Genet 7, 805–812 (2006). https://doi.org/10.1038/nrg1943
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DOI: https://doi.org/10.1038/nrg1943
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