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Efficient selection of μ −m mutants from μm-expressing myeloma cells by treatment with ricin A-conjugated anti-μ antibody

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Somatic Cell and Molecular Genetics

Abstract

We have developed an efficient system for obtaining myeloma mutants defective intrans-acting factors required for immunoglobulin (Ig) gene expression. The system consists of a myeloma cell line designed for this purpose and an efficient method for selecting mutants from it. The cell line is X63.653 transfected with the μ gene, whose tailpiece sequence was replaced with the transmembrane sequence of human EGF receptor to hold μ on the cell surface and whose CH1 sequence was removed to prevent μ from being retained in the endoplasmic reticulum. It efficiently and stably expressed μ chains of IgM on the cell surface (μ +m ) without light chains. To obtain mutants lacking μm (μ −m ) from the μ +m cell line by selectively killing μ +m cells, a method with ricin A-conjugated anti-μ antibody was more reliable than complement lysis mediated by anti-μ antibody. Applying the system, we obtained a variety of μ −m mutants.

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Literature cited

  1. Maniatis, T., Goodbourn, S., and Fisher, J.A. (1987).Science 2361237–1245.

    Google Scholar 

  2. Baeuerle, P.A. (1991).Biochim. Biophys. Acta 107263–80.

    Google Scholar 

  3. Müller, M.M., Ruppert, S., Schaffner, W., and Matthias, P. (1988).Nature 336544–551.

    Google Scholar 

  4. Scheidereit, C., Cromlish, J.A., Gerster, T., Kawakami, K., Balmaceda, C.G., Currie, R.A., and Roeder, R.G. (1988).Nature 336551–557.

    Google Scholar 

  5. Weitzman, S., and Scharff, M.D. (1976).J. Mol. Biol. 102237–252.

    Google Scholar 

  6. Shulman, M.J., Heusser, C., Filkin, C., and Köhler, G. (1982).Mol. Cell. Biol. 21033–1043.

    Google Scholar 

  7. Trimble, W.S., Baker, M.D., Boulianne, G.L., Murialdo, H., Hozumi, N., and Shulman, M.J. (1986).Somat. Cell Mol. Genet. 12467–477.

    Google Scholar 

  8. Atchison, M.L., and Perry, R.P. (1987).Cell 48121–128.

    Google Scholar 

  9. Neuberger, M.S. (1983).EMBO J. 21373–1378.

    Google Scholar 

  10. Ueda, H., Kikuchi, M., Yagi, S., and Nishimura, H. (1992).Bio/Technol. 10430–433.

    Google Scholar 

  11. Kearney, J.F., Radbruch, A., Liesegang, B., and Rajewsky, K. (1979).J. Immunol. 1231548–1550.

    Google Scholar 

  12. Lambson, B.E., Bernstein, R., and Mendelow, B.V. (1990).Somat. Cell Mol. Genet. 1691–95.

    Google Scholar 

  13. Potter, H., Weir, L., and Leder, P. (1984).Proc. Natl. Acad. Sci. U.S.A. 817161–7165.

    Google Scholar 

  14. Seon, B.K. (1984).Cancer Res. 44259–264.

    Google Scholar 

  15. Cumber, A.J., Forrester, J.A., Foxwell, B.M.J., Ross, W.C.J., and Thorpe, P.E. (1985).Methods Enzymol. 112207–225.

    Google Scholar 

  16. Behzadian, M.A., and Shimizu, N. (1985).Somat. Cell Mol. Genet. 11579–591.

    Google Scholar 

  17. Youle, R.J., and Neville, D.M., Jr. (1980).Proc. Natl. Acad. Sci. U.S.A. 775483–5486.

    Google Scholar 

  18. Haas, I.G., and Wabl, M. (1983).Nature 306387–389.

    Google Scholar 

  19. Bole, D.G., Hendershot, L.M., and Kearney, J.F. (1986).J. Cell Biol. 1021558–1566.

    Google Scholar 

  20. Hendershot, L., Bole, D., Köhler, G., and Kearney, J.F. (1987).J. Cell Biol. 104761–767.

    Google Scholar 

  21. Hombach, J., Leclercq, L., Radbruch, A., Rajewsky, K., and Reth, M. (1988).EMBO J. 73451–3456.

    Google Scholar 

  22. Hombach, J., Tsubata, T., Leclercq, L., Stappert, H., and Reth, M. (1990).Nature 343760–762.

    Google Scholar 

  23. Sitia, R., Neuberger, M.S., and Milstein, C. (1987).EMBO J. 63969–3977.

    Google Scholar 

  24. Sitia, R., Neuberger, M., Alberini, C., Bet, P., Fra, A., Valetti, C., Williams, G., and Milstein, C. (1990).Cell 60781–790.

    Google Scholar 

  25. Shaw, A.C., Mitchell, R.N., Weaver, Y.K., Campos-Torres, J., Abbas, A.K., and Leder, P. (1990).Cell 63381–392.

    Google Scholar 

  26. Köhler, G., and Shulman, M.J. (1980).Eur. J. Immunol. 10467–476.

    Google Scholar 

  27. Andrews-Wagner, R., and Silbey, C. (1983).Somat. Cell Mol. Genet. 943–54.

    Google Scholar 

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Author notes

  1. T. Shindo

    Present address: Department of Chemical Engineering, Faculty of Engineering, University of Tokyo, 7-3-1 Hongo Bunkyo-ku, 113, Tokyo, Japan

Authors and Affiliations

  1. Engineering Research Laboratories, Research Institute, Kaneka Corporation, 1-8 Miyamae, Takasago, 676, Hyogo, Japan

    T. Shindo

  2. Department of Chemical Engineering, Faculty of Engineering, University of Tokyo, 7-3-1 Hongo Bunkyo-ku, 113, Tokyo, Japan

    H. Ueda, F. Makishima, E. Suzuki & H. Nishimura

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  1. T. Shindo
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  2. H. Ueda
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  3. F. Makishima
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  4. E. Suzuki
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  5. H. Nishimura
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Shindo, T., Ueda, H., Makishima, F. et al. Efficient selection of μ −m mutants from μm-expressing myeloma cells by treatment with ricin A-conjugated anti-μ antibody. Somat Cell Mol Genet 18, 553–558 (1992). https://doi.org/10.1007/BF01232651

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  • DOI: https://doi.org/10.1007/BF01232651

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