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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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.
Bates, G.P., Wainwright, B.J., Williamson, R., and Brown, S.D.M. (1986).Mol. Cell. Biol. 63826–3830.
Weith, A., Winking, H., Brackmann, B., Boldyreff, B., and Traut, W. (1987).EMBO J. 61295–1300.
Martinsson, T., Weith, A., Cziepluch, C., and Schwab, M. (1989).Genes Chromosomes Cancer 167–78.
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.
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.
Kerem, B., Rommens, J.M., Buchanan, J., Markiewiecz, D., Cox, T.K., Chakravarti, A., Buchwald, M., and Tsui, L.C. (1989).Science 2451073–1080.
Mornet, E., Simon-Bouy, B., Serre, J.L., Estivill, X., Farrall, M., Williamson, R., Boue, J., and Boue, A. (1988).Lancet 1376–378.
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.
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.
Maniatis, T., Fritsch, E.F., and Sambrook, J. (1982).Molecular Cloning. A Laboratory Manual (Cold Spring Harbor Laboratory, Cold Spring Harbor, New York).
Fisher, E.M.C., Cavanna, J.S., and Brown, S.D.M. (1985).Proc. Natl. Acad. Sci. U.S.A. 825846–5849.
Greenfield, A.J., and Brown, S.D.M. (1987).Genomics 1153–158.
Röhme, D., Fox, H., Herrmann, B., Frischauf, A.M., Edström, J.E., Mains, P., Silver, L.M., and Lehrach, H. (1984).Cell 36783–788.
Sealy, P.G., Whittaker, P.A., Southern, E.M. (1985).Nucleic Acids Res. 131905–1922.
Lüdecke, H.-J., Senger, G., Claussen, U., and Horsthemke, B. (1989).Nature 338348–350.
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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