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BAC Resource for Molecular Cytogenetics

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Molecular Cytogenetics

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 204))

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Abstract

Bacterial artificial chromosomes (BACs) are ideal materials to use for the purpose of integrating DNA sequence with cytogenetic markers. They have been the major vectors used in genome sequencing. BACs are also well suited for fluorescent in situ hybridization (FISH) in that they represent a stable and easily manipulated form of cloned DNA that produces bright, well-defined signals on metaphase and interphase chromosome preparations (1). To link chromosomal position with DNA sequence throughout the human genome, we have developed an integrated BAC resource (1) by using FISH, PCR, and sequencing. The Resource contains a total of 6000 randomly mapped BAC clones, out of which, 1021 are BAC-sequence-tagged sites (STS) pairs representing 957 BACs (Fig. 1). This tool can be now used to rapidly identify genes affected by chromosomal rearrangements seen in genetic disorders and cancers. After this initial development, an international effort has assembled a collection of BAC clones that are both sequence-tagged and mapped relative to cytogenetic bands using FISH, resulting in a collection of 7600 clones (2; see Note 1).

An Integrated BAC Resource was developed using FISH and PCR to link chromosomal position with DNA sequence in the human genome (1). This map contains a total of 1021 BAC-STS pairs representing 957 BACs, each mapping to the position indicated by the vertical lines at the right of each chromosome ideogram. The length of the line indicates the resolution of the assignment, i.e, the outer boundaries of the band(s) within which the BAC signal is located as described in the methods. Although higher resolution may be obtained from our archived images, within a single band region, BACs are also ordered by the genetic and RH maps (1,16).

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References

  1. Korenberg J. R., Chen X.-N., Sun Z., et al. (1999) Human genome anatomy: BACs integrating the genetic and cytogenetic maps for bridging genome and biomedicine. Genome Res. 9, 994–1001.

    Article  CAS  PubMed  Google Scholar 

  2. Cheung V. G., Nowak N., Jang W., et al. (2001) Integration of cytogenetic landmarks into the draft sequence of the human genome. Nature 409, 953–958.

    Article  CAS  PubMed  Google Scholar 

  3. Bolzer A., Craig J. M., Cremer T., and Speicher M. R. (1999) A complete set of repeatdepleted, PCR-amplifiable, human chromosome-specific painting probes. Cytogenet. Cell Genet. 84, 233–240.

    Article  CAS  PubMed  Google Scholar 

  4. Lichter P. (1997) Multicolor FISHing: what’s the catch? Trends Genet. 13, 475–479.

    Article  CAS  PubMed  Google Scholar 

  5. Langer P. R., Waldrop A. A., and Ward D. C. (1981) Enzymatic synthesis of biotinlabeled polynucleotides: novel nucleic acid affinity probes. Proc. Natl. Acad. Sci. USA 11, 6633–6637.

    Article  Google Scholar 

  6. Pinkel D., Straume T., and Gray J. W. (1986) Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridization. Proc. Natl. Acad. Sci. USA 83, 2934–2938.

    Article  CAS  PubMed  Google Scholar 

  7. Johnson G. D. and Nogueira Araujo G. M. (1981) A simple method of reducing the fading of immunofluorescence during microscopy. J. Immunol. Meth. 43, 349–350.

    Article  CAS  Google Scholar 

  8. Korenberg J. R. and Chen X.-N. (1995) Human cDNA mapping using a high-resolution R-banding technique and fluorescence in situ hybridization. Cytogenet. Cell Genet. 69, 196–200.

    Article  CAS  PubMed  Google Scholar 

  9. Mitelman E. (1995) ISCN 1995 An International System for Human Cytogenetic Nomenclature.

    Google Scholar 

  10. Hubert R. S., Mitchell S., Chen X.-N., et al. (1997) BAC and PAC contigs covering 3.5 Mb of the Down syndrome congenital heart disease region between D21S55 and MX1 on chromosome 21. Genomics 41, 218–226.

    Article  CAS  PubMed  Google Scholar 

  11. Chen X.-N., Knauf J. A., Gonsky R., et al. (1998) From amplification to gene in thyroid cancer: a high-resolution mapped bacterial-artificial-chromosome resource for cancer chromosome aberrations guides gene discovery after comparative genome hybridization. Am. J. Hum. Genet. 63, 625–637.

    Article  CAS  PubMed  Google Scholar 

  12. Yamakawa K., Huo Y. K., Haendelt M. A., et al. (1998) DSCAM: a novel member of the immunoglobulin superfamily maps in a Down syndrome region and is involved in the development of the nervous system. Hum. Mol. Genet. 7, 227–237.

    Article  CAS  PubMed  Google Scholar 

  13. Korenberg J. R., Chen X.-N., Hirota H., et al. (2000) VI. Genome structure and cognitive map of Williams syndrome. J. Cogn. Neurosci. 12(Suppl), 89–107.

    Article  PubMed  Google Scholar 

  14. Chen X.-N., Mitchell S., Sun Z.-G, et al. (1996) The Integrated BAC Resource: A Powerful New Molecular Tool For Prenatal Genetics. Am. J. Hum. Genet. 59(suppl)(4), A12, Abstract #53

    Google Scholar 

  15. Knauf J. A., Elisei R., Mochly-Rosen D., et al. (1999) Involvement of protein kinase Cepsilon (PKCepsilon) in thyroid cell death. A truncated chimeric PKCepsilon cloned from a thyroid cancer cell line protects thyroid cells from apoptosis. J. Biol. Chem. 274, 23414–23425.

    Article  CAS  PubMed  Google Scholar 

  16. Hudson T. J., Stein L. D., Gerety S. S., et al. (1995) An STS-based map of the human genome. Science 270, 1945–1954.

    Article  CAS  PubMed  Google Scholar 

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© 2002 Humana Press Inc.

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Chen, XN., Korenberg, J.R. (2002). BAC Resource for Molecular Cytogenetics. In: Fan, YS. (eds) Molecular Cytogenetics. Methods in Molecular Biology™, vol 204. Humana Press. https://doi.org/10.1385/1-59259-300-3:391

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  • DOI: https://doi.org/10.1385/1-59259-300-3:391

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-006-9

  • Online ISBN: 978-1-59259-300-2

  • eBook Packages: Springer Protocols

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