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Localization in mouse-L-cell chromosomal sites of transferred immunoglobulin genes

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Abstract

Mouse thymidine kinase negative (tk) L-cells were cotransformed with two different κ light chain genes (cloned from mouse myeloma) and the tk gene from Herpes simplex virus I. (Transformation is defined as change in the genotype by introduction of foreign DNA.) About 80% of the tk +-transformants contained varying amounts of transferred κ light chain sequences, one transformant about 150 copies per genome. The transferred immunoglobulin genes appear to be organized in a nucleosomal substructure, as deduced from digestion experiments with micrococcal nuclease. In situ hybridization experiments revealed, that the transferred genes are not distributed randomly across the chromosomes of the recipient cell. Instead they are clustered at one or a few chromosomal locations.

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References

  • Anderson, W.F., Killos, L., Sanders-Haigh, L., Kretschmer, P.J., Diacumacos, E.G.: Replication and expression of thymidine kinase and human globin genes microinjected into mouse fibroblasts. Proc. nat. Acad. Sci. (Wash.) 77, 5399–5403 (1980)

    Google Scholar 

  • Camerini-Otero, R.D., Zaslow, M.A.: Nucleosomal packaging of the thymidine kinase gene of herpes simplex virus transferred into mouse cells: An actively expressed single copy gene. Proc. nat. Acad. Sci (Wash.) 77, 5079–5083 (1980)

    Google Scholar 

  • Capecchi, M.R.: High efficiency transformation by direct microinjection of DNA into mammalian cells. Cell 22, 479–488 (1980)

    Google Scholar 

  • Fittler, F., Zachau, H.G.: Subunit structure of α-satellite DNA containing chromatin from African green monkey cells. Nuclei Acids Res. 7, 1–13 (1979)

    Google Scholar 

  • Graham, F.L., Eb, A.J. van der: The new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52, 456–467 (1973)

    Google Scholar 

  • Graham, F.L., Bacchetti, S., McKinnon, R., Stanners, C., Cordell, B., Goodmann, H.: Introduction of macromolecules into viable mammalian cells (R. Baserga, C. Croce and G. Rovera, eds.). Wistar Symp. workshop. New York: Alan R. Riss Inc. 1979

    Google Scholar 

  • Henderson, A.S., Yu, M.T., Silverstein, S.: Chromosomal DNA homologous to herpes simplex virus I in a mouse L-cell line. Cytogenet. Cell Genet. 29, 107–115 (1981)

    Google Scholar 

  • Jeffreys, A.J., Flavell, R.A.: A physical map of the DNA regions flanking the rabbit β-globulin gene. Cell 12, 429–439 (1977)

    Google Scholar 

  • Lewis, W.H., Srinivasan, P.R., Stokoe, N., Siminovitch, L.: Parameters governing the transfer of the genes for thymidine kinase and dihydrofolate reductase into mouse cells using metaphase chromosomes or DNA. Somat. Cell Genet. 6, 333–347 (1980)

    Google Scholar 

  • Littlefield, J.W.: Selection of hybrids from matings of fibroblasts in vitro and their presumed recombinants. Science 145, 709 (1964)

    Google Scholar 

  • Maio, J.J., Schildkraut, C.L.: Isolated mammalian metaphase chromosomes, I. General characteristics of nucleic acids and proteins. J. molec. Biol. 24, 29–39 (1967)

    Google Scholar 

  • Mantei, N., Boll, W., Weissmann, Ch.: Rabbit β-globin mRNA production in mouse L-cells transformed with cloned rabbit β-globin chromosomal DNA. Nature (Lond.) 281, 40–46 (1979)

    Google Scholar 

  • Müller, C., Graessmann, A., Graessmann, M.: Mapping of early SV 40-specific functions by microinjection of different early viral DNA fragments. Cell 15, 579–585 (1978)

    Google Scholar 

  • Pellicer, A., Robins, D., Wold, B., Sweet, R., Jackson, J., Lowy, I., Roberts, J.M., Sim, G.K., Silverstein, S., Axel, R.: Altering genotype and phenotype by DNA-mediated gene transfer. Science 209, 1414–1422 (1980)

    Google Scholar 

  • Perucho, M., Hanahan, D., Wigler, M.: Genetic and physical linkage of exogenous sequences in transformed cells. Cell 22, 309–317 (1980)

    Google Scholar 

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

    Google Scholar 

  • Robins, D.M., Ripley, S., Henderson, A.S., Axel, R.: Transforming DNA integrates into the host chromosome. Cell 23, 29–39 (1981)

    Google Scholar 

  • Shapiro, J.M., Moar, M.H., Ohno, S., Klein, G.: Acetic acid treatment denatures DNA while preserving chromosomal morphology during the in situ hybridization procedure. Exp. Cell Res. 115, 411–414 (1978)

    Google Scholar 

  • Southern, E.M.: Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. molec. Biol. 98, 503–517 (1975)

    Google Scholar 

  • Steffen, D., Weinberg, R.A.: The integrated genome of murine leukemia virus. Cell 15, 1003–1010 (1978)

    Google Scholar 

  • Steinmetz, M., Zachau, H.G., Mach, B.: Cloning of immunoglobulin kappa light chain genes from mouse liver and myeloma MOPC 173. Nucleic Acids Res. 6, 3213–3230 (1979)

    Google Scholar 

  • Steinmetz, M., Zachau, H.G.: Two rearranged immunoglobulin kappa light chain genes in one mouse myeloma. Nucleic Acids Res. 8, 1693–1707 (1980)

    Google Scholar 

  • Weissmann, Ch., Mantei, N., Boll, W., Weaver, R.F., Wilkie, N., Clements, B., Taniguchi, T., Ooyen, A. van, Berg, J. van den, Fried, M., Murray, K.: From the gene to protein: Information transfer in normal and abnormal cells. Proc. Miami Winter Symp. (Russel, T.R., Brew, K., Faber, H. and Schultz, J., eds.) vol. 16, pp. 99–132. New York: Acad. Press

  • Wigler, M., Silverstein, S., Lee. L.S., Pellicer, A., Cheng, Y., Axel, R.: Transfer of purified herpes virus thymidine kinase gene to cultured mouse cells. Cell 11, 223–232 (1977)

    Google Scholar 

  • Wigler, M., Sweet, R., Sim, G.K., Wold, B., Pellicer, A., Lacy, E., Maniatis, T., Silverstein, S., Axel, R.: Transformation of mammalian cells with genes from procaryotes and eucaryotes. Cell 16, 777–785 (1979a)

    Google Scholar 

  • Wigler, M., Pellicer, A., Silverstein, S., Axel, R., Urlaub, G., Chasin, L.: DNA mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells. Proc. nat. Acad. Sci. (Wash.) 76, 1373–1376 (1979b)

    Google Scholar 

  • Willecke, K., Ruddle, F.H.: Transfer of the human gene for hypoxanthine-guanine phosphoribosyl-transferase via isolated human metaphase chromosomes into mouse L-cells. Proc. nat. Acad. Sci. (Wash.) 72, 1792–1796 (1975)

    Google Scholar 

  • Wong, K.T., Nicolau, C., Hofschneider, H.P.: Appearance of β-lactamase activity in animal cells upon liposome-mediated gene transfer. Gene 10, 87–94 (1980)

    Google Scholar 

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Fittler, F., Bogenberger, J. & Steinmetz, M. Localization in mouse-L-cell chromosomal sites of transferred immunoglobulin genes. Chromosoma 84, 717–727 (1982). https://doi.org/10.1007/BF00286336

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