Chromosome Painting Using Chromosome-Specific BAC Clones
Language English Country United States Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
- Keywords
- Arabidopsis thaliana, BAC FISH, Brassicaceae, Chromosome painting, Fluorescence in situ hybridization (FISH), Nick translation,
- MeSH
- Arabidopsis * genetics MeSH
- Brassicaceae * genetics MeSH
- Clone Cells MeSH
- Chromosomes MeSH
- DNA Probes MeSH
- DNA MeSH
- In Situ Hybridization, Fluorescence methods MeSH
- Chromosome Painting methods MeSH
- Chromosomes, Artificial, Bacterial genetics MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- DNA Probes MeSH
- DNA MeSH
Chromosome painting (CP) refers to visualization of large chromosome regions, chromosome arms or entire chromosomes via fluorescence in situ hybridization (FISH) of chromosome-specific DNA sequences. For CP in crucifers (Brassicaceae), typically contigs of chromosome-specific bacterial artificial chromosomes (BAC) from Arabidopsis thaliana are applied as painting probes on chromosomes of A. thaliana or other species (comparative chromosome painting, CCP). CP/CCP enables to identify and trace particular chromosome regions and/or chromosomes throughout all mitotic and meiotic stages as well as corresponding interphase chromosome territories. However, extended pachytene chromosomes provide the highest resolution of CP/CCP. Fine-scale chromosome structure, structural chromosome rearrangements (such as inversions, translocations, centromere repositioning), and chromosome breakpoints can be investigated by CP/CCP. BAC DNA probes can be accompanied by other types of DNA probes, such as repetitive DNA, genomic DNA, or synthetic oligonucleotide probes. Here, we describe a robust step-by-step protocol of CP and CCP which proved to be efficient across the family Brassicaceae, but which is also applicable to other angiosperm families.
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