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Gene mapping and physical arrangements of human chromatin in transformed, hybrid cells: Fluorescent and autoradiographic in situ hybridization compared

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Somatic Cell and Molecular Genetics

Abstract

We compare a fluorescent in situ hybridization technique, usingN-acetoxy-2-acetylaminofluorene (N-ACO-AAF) modified DNA adducts, with3H-labeled DNA in situ hybridization for (1) visualizing human transgenomes in HRAS1-selected, chromosome-mediated gene transfer (CMGT), and (2) mapping chromosomal SV40 in an SV40-transformed, human-mouse hybrid cell line. We demonstrate that individual HRAS1-CMGTs may contain multiple fragments of human chromatin. We deduce that the CMGT process can involve interstitial loss of mouse chromatin. We conclude that the N-ACO-AAF technique gives finer resolution than3H-labeled in situ hybridization. However,3H-labeling is more sensitive and has allowed us to sublocalize SV40 in Cl21 to the region 7q31–35.

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Author notes

  1. A. R. Mitchell

    Present address: 3216 Gordon H. Scott Hall of Basic Medical Sciences, Wayne State University, 540 East Canfield Avenue, 48201, Detroit, Michigan

  2. J. E. N. Morten

    Present address: Paterson Laboratories, Christie Hospital and Holt Radium Institute, Wilmslow Road, M2O 9BX, Manchester, UK

Authors and Affiliations

  1. MRC Clinical and Population Cytogenetics Unit, Western General Hospital, Crewe Road, EH4 2XU, Edinburgh, UK

    A. R. Mitchell, J. R. Gosden, J. E. N. Morten & D. J. Porteous

  2. Institute of Molecular Biology, Austrian Academy of Science, Billrothstrusse II, A-5020, Salzburg

    P. Ambros

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  1. A. R. Mitchell
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  2. P. Ambros
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  4. J. E. N. Morten
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  5. D. J. Porteous
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Mitchell, A.R., Ambros, P., Gosden, J.R. et al. Gene mapping and physical arrangements of human chromatin in transformed, hybrid cells: Fluorescent and autoradiographic in situ hybridization compared. Somat Cell Mol Genet 12, 313–324 (1986). https://doi.org/10.1007/BF01570725

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  • DOI: https://doi.org/10.1007/BF01570725

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