期刊
CYTOGENETIC AND GENOME RESEARCH
卷 107, 期 1-2, 页码 14-17出版社
KARGER
DOI: 10.1159/000079565
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资金
- NATIONAL CANCER INSTITUTE [R01CA043322] Funding Source: NIH RePORTER
- NCI NIH HHS [CA-43322] Funding Source: Medline
- PHS HHS [76260] Funding Source: Medline
The ability to prepare single-stranded chromosomal target DNA allows innovative uses of FISH technology for studies of chromosome organization. Standard FISH methodologies require functionally single-stranded DNAs in order to facilitate hybridization between the probe and the complementary chromosomal target sequence. This usually involves denaturation of double-stranded probes to induce temporary separation of the DNA strands. Strand-specific FISH (CO-FISH; Chromosome Orientation-FISH) involves selective removal of newly replicated strands from DNA of metaphase chromosomes which results in single-stranded target DNA. When single-stranded probes are then hybridized to such targets, the resulting strand-specific hybridization is capable of revealing a level of information previously unattainable at the cytogenetic level. Mammalian telomeric DNA consists of tandem repeats of the (TTAGGG) sequence, oriented 5'-->3' towards the termini of all vertebrate chromosomes. Based on this conserved structural organization, CO-FISH with a telomere probe reveals the absolute 5'-->3' orientation of DNA sequences with respect to the pter-->qter direction of chromosomes. Development and various applications of CO-FISH will be discussed: detection of cryptic inversions, discrimination between telomeres produced by leading-versus lagging-strand synthesis, and replication timing of mammalian telomeres. Copyright (C) 2004 S. Karger AG, Basel.
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