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Molecular analysis of human Chromosome 16 cosmid clones containing NotI sites

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Abstract

To test the feasibility of using cloned NotI sites as markers for physical mapping, we have screened for cosmid clones spanning the NotI sites on human Chromosome (Chr) 16. Fluorescence in situ hybridization analysis of these clones confirms the previously reported cluster of NotI sites on 16p13.3. Methylation status of the cloned NotI sites on genomic DNA was established by hybridization of the cosmids to Southern blots containing EcoRI and EcoRI/NotI digest of genomic DNA. These results indicated that four of six clones included in our study can be used as linking clones for physical mapping. Two clones have NotI sites which are not cleavable in the cell lines tested. In one clone, the NotI site exists as an isolated rare-cutting restriction enzyme site, whereas in the other clone the NotI site appears to be island-related.

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References

  • Anand, R.: Pulsed field gel electrophoresis: A technique for fractionating large DNA molecules. Trends Genet 278–283, 1986.

  • Barlow, D.P. and Lehrach, H.: Genetics by gel electrophoresis: The impact of pulsed field gel electrophoresis on mammalian genetics. Trends Genet 3: 167–171, 1987.

    Google Scholar 

  • Bates, P.F.: Double cos site vectors: simplified cosmid cloning. Methods Enzymol 153: 82–94, 1987.

    Google Scholar 

  • Bates, P.F., and Swift, R.A.: Double cos site vectors: Simplified cosmid cloning. Gene 26: 137–146, 1983.

    Google Scholar 

  • Breuning, M.H., Medan, K., Verjaal, M., Wijnen, J.T., Meera Klou, P., and Pearson, P.L.: Human α-globin maps to pter-p 13.3 in chromosome 16 distal to PGP. Hum Genet 76: 287–289, 1987.

    Google Scholar 

  • Brown, W.R.A. and Bird, A.P: Long-range restriction site mapping of mammalian genomic DNA. Nature 322: 477–481, 1986.

    Google Scholar 

  • Buiting, K., Passarge, E., and Horsthemke, B.: Construction of a chromosome 15-specific linking library and identification of potential gene sequences. Genomics 3: 143–149, 1988.

    Google Scholar 

  • Callen, D.F.: A mouse-human hybrid cell panel for mapping human chromosome 16. Ann Genet 29: 235–239, 1986.

    Google Scholar 

  • Callen, D.F., Hyland, V.J., Baker, E.G. Fratini, A., Simmers, R.N., Mulley, J.C., and Sutherland, G.R.: Fine mapping of gene probes and anonymous DNA fragments to the long arm of chromosome 16. Genomics 2: 144–153, 1988.

    Google Scholar 

  • Callen, D.F., Hyland, V.J., Baker, E.G., Fratini, A., Gedeon, A.K., Mulley, J.C., Fernandez, K.E.W., Breuning, M.H., and Sutherland, G.R.: Mapping the short arm of chromosome 16. Genomics 4: 348–354, 1989.

    Google Scholar 

  • Cami, B. and Kourilsky, P.: Screening of cloned recombinant DNA in bacteria by in situ hybridization. Nucl Acids Res 5: 2381–2390, 1978.

    Google Scholar 

  • Campbell, D.A.: c2X75, a derivative of the cosmid vector c2XB. Nucl Acids Res 17: 458, 1989.

    Google Scholar 

  • Cedar, H.: DNA methylation and gene activity. Cell 53: 3–4, 1988.

    Google Scholar 

  • Chu, G., Vollrath, D., and Davis, R.R.: Separation of large DNA molecules by contour-clamped homogeneous electric fields. Science 232: 1582–1585, 1986.

    Google Scholar 

  • Church, G.M. and Gilbert, W.: Genomic sequencing. Proc Natl Acad Sci USA 81: 1991–1995, 1984.

    Google Scholar 

  • Dilella, A.G. and Woo, S.L.C.: Cloning large segments of genomic DNA using cosmid vectors. Methods Enzymol 152: 199–212, 1987.

    Google Scholar 

  • Evans, G.A., and Wahl, G.M.: Cosmid vectors for genomic walking and rapid restriction mapping. Methods Enzymol 152: 604–610, 1987.

    Google Scholar 

  • Evans, G.A., Lewis, K., and Rothenberg, B.E.: High efficiency vectors for cosmid microcloning and genomic analysis. Gene 79: 9–20, 1989.

    Google Scholar 

  • Feinberg, A. and Vogelstein, B.: A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13, 1983.

    Google Scholar 

  • Feinberg, A. and Vogelstein, B.: Addendum. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 137: 266–277, 1984.

    Google Scholar 

  • Fischel-Ghodsian, N., Nicholls, R.D., and Higgs, D.R.: Unusual features of CpG-rich HTF islands in the human α-globin complex. Nucl Acids Res 15: 9215–9225, 1987.

    Google Scholar 

  • Harris, P.C., Barton, N.M., Higgs, D.R., Reeders, S.T., and Wilkie, A.O.M.: A long-range restriction map between the α-globin complex and a marker closely linked to the polycystic kidney disease I (PKDI) locus. Genomics 7: 195–206, 1990.

    Google Scholar 

  • Hermann, B.G., Barlow, D.P., and Lehrach, H.: A large inverted duplication allows homologous recombination between chromosomes heterozygous for the proximal T complex inversion. Cell 48: 813–825, 1987.

    Google Scholar 

  • Himmelbauer, H., Germino, G.G., Ceccherini, I., Romeo, G., Reeders, S.T., and Frischauf, A.M.: Saturating the region of the polycystic kidney disease gene with NotI linking clones. Am J Hum Genet 48: 325–334, 1991.

    Google Scholar 

  • Holmes, D.S. and Quigley, M.: A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114: 193–197, 1981.

    Google Scholar 

  • Ito, T. and Sakaki, Y.: A novel procedure for selective cloning of NotI linking fragments from mammalian genomes. Nucl Acids Res 16: 9177–9184, 1988.

    Google Scholar 

  • Koeffler, H.P., Sparkers, R.S., Stang, H., and Mohandas T.: Regional assignment of genes for human α-globin and phosphoglycolate phosphatase to the short arm of chromosome 16. Proc Natl Acad Sci USA 78: 7015–7018, 1981.

    Google Scholar 

  • Korenberg, J.R. and Rykowski, M.C.: Human genome organization: Alu, lines, and the molecular structure of metaphase chromosome bands. Cell 53: 391–400, 1988.

    Google Scholar 

  • Lerner, T.J., Kessler, D., Landes, G., Bird, J., Leverone, B., Watkins, P.A., and Klinger, K.: Identification of cosmids containing rare restriction sites by FIGE. Cytogenet Cell Genet 46: 649, 1987.

    Google Scholar 

  • Lichter, P., Tang, C-J.C., Call, K., Hermanson, G., Evans, G.A., Housman, D., and Ward, D.: High resolution mapping of human chromosome 11 by in situ hybridization with cosmid clones. Science 247: 64–69, 1990.

    Google Scholar 

  • Lindsay, S. and Bird, A.: Use of restriction enzymes to detect potential gene sequences in mammalian DNA. Nature 327: 336–338, 1987.

    Google Scholar 

  • Lindsay, S., Adams, R.L.P., and Monk, M.: Til1—a human lymphoblastoid cell line with minimal DNA methylation. Hum Genet 81: 252–256, 1989.

    Google Scholar 

  • Litt, M. and White, R.L.: A highly polymorphic locus in human DNA revealed by cosmid-derived probes. Proc Natl Acad Sci USA 82: 6206–6210, 1985.

    Google Scholar 

  • Pohl, T.M., Zimmer, M., MacDonald, M.E., Smith, B., Bucan, M., Poustka, A., Volina, S., Searle, S., Zehetner, G., Wasmuth, J.J., Gusella, J., Lehrach, H., and Frischchauf, A.-M: Construction of a NotI linking library and isolation of new markers close to the Huntington's disease gene. Nucl Acids Res 16: 9185–9198, 1988.

    Google Scholar 

  • Poustka, A. and Lehrach, H: Jumping libraries and linking libraries: The next generation of molecular tools in mammalian genetics. Trends Genet 2: 174–179, 1986.

    Google Scholar 

  • Rappold, G.A. and Lehrach, H.: A long range restriction map of the pseudoautosomal region by partial digest PFGE analysis from the telomere. Nucl Acids Res 16: 5361–5377, 1988.

    Google Scholar 

  • Rigby, P.W.J., Dieckmann, M., Rhodes, C., and Berg, P.: Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113: 237–251, 1977.

    Google Scholar 

  • Sealy, P.G., Whittaker, P.A., and Southern, E.M.: Removal of repeated sequences from hybridization probes. Nucl Acids Res 13: 1905–1922, 1985.

    Google Scholar 

  • Smith, D.I., Golembieski, W., Gilbert, J.D., Kzyma, L., and Miller, O.J.: Overabundance of rare-cutting restriction endonuclease sites in the human genome. Nucl Acids Res 15: 1173–1184, 1987.

    Google Scholar 

  • Smith, G.E. and Summers, M.D.: The bidirectional transfer of DNA and RNA to nitrocellulose or diazobenzyloxymethyl-paper. Anal Biochem 109: 123–129, 1980.

    Google Scholar 

  • Smith, H.O. and Birnstiel, M.L.: A simple method for DNA restriction site mapping. Nucl Acids Res 3: 2387–2398, 1976.

    Google Scholar 

  • Trasler, J.M., Hake, L.E., Johnson, P.A., Alcivar, A.A., Milette, C.F., and Hecht, N.B.: DNA methylation and demethylation events during meiotic prophase in the mouse testis. Mol Cell Biol 10: 1828–1834, 1990.

    Google Scholar 

  • Tratner, I., Nahon, J.L. and Sala-Trepat, J.M.: Differences in methylation patterns of the α-fetoprotein and albumin genes in hepatic and non-hepatic developing rat tissues. Nucl Acids Res 16: 2749–2763, 1988.

    Google Scholar 

  • Wahl, G.M., Lewis, K.A., Ruiz, J.C., Rothenberg, B., Zhao, J., and Evans, G.A.: Cosmid vectors for genomic walking, restriction mapping, and gene transfer. Proc Natl Acad Sci USA 84: 2160–2164, 1987.

    Google Scholar 

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Lerner, T., Wright, G., Leverone, B. et al. Molecular analysis of human Chromosome 16 cosmid clones containing NotI sites. Mammalian Genome 3, 92–100 (1992). https://doi.org/10.1007/BF00431252

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