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Microdissection and microcloning of human chromosome 7q22-32 region

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Somatic Cell and Molecular Genetics

Abstract

Genetic information contributing to cystic fibrosis in addition on the CF gene is suggested to reside on the long arm of the human chromosome 7. In our attempt to analyze this genomic region in detail, we generated a region-specific DNA probe library by microdissection and microcloning of the midpiece of the chromosome 7q arm. Microdissection was performed in unstained metaphase spreads from a human × mouse hybrid cell line containing chromosome 7 as the only human chromosome. We obtained 593 clones from 75 dissected chromosomal fragments. At least 88% of the microclones were true recombinants; 40% of the clones contained repetitive sequences as determined by plaque hybridization with genomic DNA as probe. The overall mean fragment size of insert fragments was 3.2 kb, the median size was 3.5 kb. Regional mapping of 30 DNA fragments was performed by the aid of hybrid cell lines containing different segments of human chromosome 7; 50% of the microcloned inserts were found to map to 7q22-32.

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Literature cited

  1. Kaiser, R., Weber, J., Grzeschik, K.-H., Edström, J.E., Driesel, A., Zengerling, S., Buchwald, M., Tsui, L.C., and Olek, K. (1987).Mol. Biol. Rep. 123–6.

    PubMed  Google Scholar 

  2. Bates, G.P., Wainwright, B.J., Williamson, R., and Brown, S.D.M. (1986).Mol. Cell. Biol. 63826–3830.

    PubMed  Google Scholar 

  3. Weith, A., Winking, H., Brackmann, B., Boldyreff, B., and Traut, W. (1987).EMBO J. 61295–1300.

    PubMed  Google Scholar 

  4. Martinsson, T., Weith, A., Cziepluch, C., and Schwab, M. (1989).Genes Chromosomes Cancer 167–78.

    PubMed  Google Scholar 

  5. Rommens, J.M., Iannuzzi, M.C., Kerem, B., Drumm, M.L., Melmer, G., Dean, M., Rozmahel, R., Cole, J.L., Kennedy, D., Hidaka, N., Zsiga, M., Buchwald, M., Riordan, J.R., Tsui, L.C., and Collins, F.S. (1989).Science 2451059–1065.

    PubMed  Google Scholar 

  6. Riordan, J.R., Rommens, J.M., Kerem, B., Alon, N., Rozmahel, R., Grzelczak, Z., Zielenski, J., Lok, S., Plavsic, N., Chou, J.L., Drumm, L., Iannuzzi, M.C., Collins, F.S., and Tsui, L.C. (1989).Science 2451066–1073.

    PubMed  Google Scholar 

  7. Kerem, B., Rommens, J.M., Buchanan, J., Markiewiecz, D., Cox, T.K., Chakravarti, A., Buchwald, M., and Tsui, L.C. (1989).Science 2451073–1080.

    PubMed  Google Scholar 

  8. Mornet, E., Simon-Bouy, B., Serre, J.L., Estivill, X., Farrall, M., Williamson, R., Boue, J., and Boue, A. (1988).Lancet 1376–378.

    PubMed  Google Scholar 

  9. Kerem, B., Buchanan, J.A., Durie, P., Corey, L., Levison, H., Rommens, J.M., Buchwald, M., and Tsui, L.C. (1989).Am. J. Hum. Genet. 44827.

    PubMed  Google Scholar 

  10. Pirotta, V., Jäckle, H., and Edström, J.E. (1983). InGenetic Engineering: Principles and Methods, Vol. 5 (eds.) Setlow, J.K., and Hollaender, A. (Plenum Press, New York), pp. 1–17.

    Google Scholar 

  11. Maniatis, T., Fritsch, E.F., and Sambrook, J. (1982).Molecular Cloning. A Laboratory Manual (Cold Spring Harbor Laboratory, Cold Spring Harbor, New York).

    Google Scholar 

  12. Fisher, E.M.C., Cavanna, J.S., and Brown, S.D.M. (1985).Proc. Natl. Acad. Sci. U.S.A. 825846–5849.

    PubMed  Google Scholar 

  13. Greenfield, A.J., and Brown, S.D.M. (1987).Genomics 1153–158.

    PubMed  Google Scholar 

  14. Röhme, D., Fox, H., Herrmann, B., Frischauf, A.M., Edström, J.E., Mains, P., Silver, L.M., and Lehrach, H. (1984).Cell 36783–788.

    PubMed  Google Scholar 

  15. Sealy, P.G., Whittaker, P.A., Southern, E.M. (1985).Nucleic Acids Res. 131905–1922.

    PubMed  Google Scholar 

  16. Lüdecke, H.-J., Senger, G., Claussen, U., and Horsthemke, B. (1989).Nature 338348–350.

    PubMed  Google Scholar 

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Weber, J., Weith, A., Kaiser, R. et al. Microdissection and microcloning of human chromosome 7q22-32 region. Somat Cell Mol Genet 16, 123–128 (1990). https://doi.org/10.1007/BF01233042

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  • DOI: https://doi.org/10.1007/BF01233042

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