Abstract
Transgenic mice were generated by coinjection of a dominant marker gene that induces fur and eye pigmentation (a tyrosinase minigene) plus an unrelated DNA construction that has a γ-glutamyl transferase (γGT) promoter linked to aras oncogene. Mice transgenic for γGT-ras could be identified in the first and all subsequent generations by simple visual inspection for pigmentation. Furthermore, the γ-glutamyl transferase promoter was active in kidney but not skin of the transgenic mice, indicating that the cointegrated DNA was active and independently expressed. These results confirm that the tyrosinase minigene can be used for coinjections to allow rapid visual identification of transgenic mice.
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Behringer, R.R., Ryan, T.M., Reilly, M.P., Asakura, T., Palmiter, R.D., Brinster, R.L. and Townes, T.M. (1989) Synthesis of functional human hemoglobin in transgenic mice.Science 245, 971–3.
Behringer, R.R., Ryan, T.M., Palmiter, R.D., Brinster, R.L. and Townes, T.M. (1990) Human γ- to β-globin gene switching in transgenic mice.Genes Devel. 4, 380–9.
Beermann, F., Ruppert, S., Hummler, E., Bosch, F.X., Müller, G., Rüther, U. and Schütz, G. (1990) Rescue of the albino phenotype by introduction of a functional tyrosinase gene into mice.EMBO J. 9, 2819–26.
Beermann, F., Rupper, S., Hummler, E. and Schütz, G. (1991) Tyrosinase as a marker for transgenic mice.Nucl. Acids Res. 19, 958.
Bishop, J.O. and Smith, P. (1989) Mechanism of chromosomal integration of microinjected DNA.Mol. Biol. Med. 6, 283–98.
Brinster, R.L., Chen, H.Y., Trumbauer, M., Senear, A.W., Warren, R. and Palmiter, R.D. (1981) Somatic expression of herpes thymidine kinase in mice following injection of a fusion gene into eggs.Cell,27, 223–31.
Capon, D.J., Chen, E.Y., Levinson, A.D., Seeburg, P.H. and Goeddel, D.V. (1983) Complete nucleotide sequences of the T24 human bladder carcinoma oncogene and its normal homologue.Nature 302, 33–7.
Chomczynski, P. and Sacchi, N. (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenolchloroform extraction.Anal. Biochem. 162, 156–9.
Einat, P., Bergman, Y., Yaffe, D. and Shani, M. (1987) Expression in transgenic mice of two genes of different tissue specificity integrated into a single chromosomal site.Genes Devel. 1, 1075–84.
Goodnow, C.C., Crosbie, J., Adelstein, S., Lavoie, T.B., Smith-Till, S.J., Brink, R.A., Pritchard-Briscoe, H., Wotherspoon, J.S., Loblay, R.H., Raphael K., Trent, R.J. and Basten, A. (1988) Altered immunoglobulin expression and functional silencing of self-reactive B lymphocytes in transgenic mice.Nature 334, 676–82.
Hogan, B., Costantini, F. and Lacy, E. (1986)Manipulating the Mouse Embryo. New York, USA: Cold Spring Harbor Laboratory.
Kawasaki, E.S. (1990) Amplification of RNA. In Innis, M.A., Gelfand, D.H., Sninsky, J.J. and White, T.J. eds,PCR Protocols, pp. 21–7. San Diego, USA: Academic Press.
Miller, S.P., Arya D.V. and Srivastava, S.K. (1976) Studies of γ-glutamyl transpeptidase in human ocular tissues.Exp. Eye Res. 22, 329–34.
Rajagopalan, S., Park, J.-H., Patel, P.D., Lebovitz, R.M. and Lieberman, M.W. (1990) Cloning and analysis of the rat γ-glutamyltransferase gene.J. Biol. Chem. 265, 11721–5.
Rathbun, W.B. and Wicker, K. (1973) Bovine lens γ-glutamyl transpeptidase.Exp. Eye Res. 15, 161–71.
Reynolds, V.L., Lebovitz, R.M., Warren, S., Hawley, T.S., Godwin, A.K. and Lieberman, M.W. (1987) Regulation of a metallothionein-ras T24 fusion gene by zinc results in graded alterations in cell morphology and growth.Oncogene 1, 323–30.
Rugh, R. (1968)The Mouse: Its Reproduction and Development, pp. 245. Minneapolis, USA: Burgess Publishing.
Ryan, T.M., Townes, T.M., Reilly, M.P., Asakura, T., Palmiter, R.D., Brinster, R.L. and Behringer, R.R. (1990) Human sickle hemoglobin in transgenic mice.Science 247, 566–8.
Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989)Molecular Cloning: A Laboratory Manual, 2nd Edn. New York, USA; Cold Spring Harbor Laboratory.
Southern, P.J. and Berg, P. (1982) Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter.J. Mol. Appl. Genet. 1, 327–41.
Storb, U., Pinkert, C., Arp, B., Engler, P., Gollahon, K., Manz, J., Brady, W. and Brinster, R.L. (1986) Transgenic mice with μ and κ genes encoding antiphosphorylcholine antibodies.J. Exp. Med. 164, 627–41.
Taketo, M., Schroeder, A.C., Mobraaten, L.E., Gunning, K.B., Hanten, G., Fox, R.R., Roderick, T.H., Stewart, C.L., Lilly, F., Hansen, C.T. and Overbeek, P.A. (1991) FVB/N: an inbred mouse strain preferable for transgenic analyses.Proc. Natl Acad. Sci. USA 88, 2065–9.
Tanaka, S., Yamamoto, H., Takeuchi, S. and Takeuchi, T. (1990) Melanization in albino mice transformed by introducing cloned mouse tyrosinase gene.Development 108, 223–7.
Taparowsky, E., Suard, Y., Fasano, O., Shimizu, K., Goldfarb, M. and Wigler, M. (1982) Activation of the T24 bladder carcinoma transforming gene is linked to a single amino acid change.Nature 300, 762–5.
Theiler, K. (1989)The House Mouse: Atlas of Embryonic Development, pp. 89. New York, USA: Springer-Verlag.
Yokoyama, T., Silversides, D.W., Waymire, K.G., Kwon, B.S., Takeuchi, T., Overbeek, P.A. (1990) Conserved cysteine to serine mutation in tyrosinase is responsible for the classical albino mutation in laboratory mice.Nucl. Acids Res. 18, 7293–8.
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Overbeek, P.A., Aguilar-Cordova, E., Hanten, G. et al. Coinjection strategy for visual identification of transgenic mice. Transgenic Research 1, 31–37 (1991). https://doi.org/10.1007/BF02512994
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DOI: https://doi.org/10.1007/BF02512994