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Haplotype-Specific Alphoid Oligonucleotides Can Distinguish Normal Homologous Chromosomes by Fish

Published online by Cambridge University Press:  02 July 2020

A. Gregory Matera  and
Christine L. O'Keefe
A. Gregory Matera
Affiliation:
Department of Genetics, Case Western Reserve University, School of Medicine, ClevelandOH, 44106-4955
Christine L. O'Keefe
Affiliation:
Department of Genetics, Case Western Reserve University, School of Medicine, ClevelandOH, 44106-4955
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Extract

The field of molecular cytogenetics is presently hampered by the need for allele- and homologue-specific hybridization probes. Vast stretches of the human genome display a considerable amount of polymorphic variation between individuals. Alpha satellite DNA is a tandemly repeated sequence located at the centromeres of all primate chromosomes and is a rich source of polymorphisms. These DNA variants are phenotypically silent; they are also stable and heritable in Mendelian fashion. We have shown that we can use this genetic diversity to create homologue-specific probes using fluorescence in situ hybridization (FISH) However unlike most classical heteromorphisms, which are detected by banding techniques, our approach does not depend upon detection of gross alterations in chromosome structure, but on the basis of sequence composition. We have constructed highly-specific oligonucleotide probes that are the first reported FISH probes to discriminate between the cytogenetically indistinguishable chromosomes of normal individuals.

These sequence-specific probes have been used to follow the transmission of a single chromosome 17 through three generation families, similar to a typical polymorphic marker.

Type
Genomes, Chromsomes and Genes: Emerging Technologies
Information
Microscopy and Microanalysis , Volume 3 , Issue S2: Proceedings: Microscopy & Microanalysis '97, Microscopy Society of America 55th Annual Meeting, Microbeam Analysis Society 31st Annual Meeting, Histochemical Society 48th Annual Meeting, Cleveland, Ohio, August 10-14, 1997 , August 1997 , pp. 197 - 198
Copyright
Copyright © Microscopy Society of America 1997

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References

1.C.L., O'Keefe, P.E., Warburton and A.G., Matera (1996). Oligonucleotide probes for alpha satellite DNA variants can distinguish homologous chromosomes by FISH. Human Molecular Genetics 5:1793–1799.Google Scholar
2.C.L., O'Keefe, D.K., Griffin, C.J., Bean, A.G., Matera and T.J., Hassold (1997). Alphoid variant-specific FISH probes can distinguish autosomal meiosis I from meiosis II non-disjunction in human sperm, (submitted).10.1007/s004390050587Google Scholar
3.M.A., Abruzzo and T.J., Hassold (1995). Etiology of nondisjunction in humans. Environmental and Molecular Mutagenesis 25: 38–47.Google Scholar
4.The authors acknowledge the generous support of the March of Dimes Birth Defects Foundation (Basil O'Connor Award #5-FY-96-0554) for their support of this work.Google Scholar