5780447275

914

B. Rihn et al.

It is now possible to use molecular biology techniąues with non-radioactive DNA labelling and detection, and our work on transgenic mice demonstrates their feasability and relevance for toxicological studies.

REFERENCES

E

e..______

O U) in .

<

i

Fig. 5 Evaluation ofthe transgene copy-number on a colored biot using a scanner densitometer. A) Hybridized biot of the v-Ha-ras DNA dilutions (upper) and c-Ha-ras DNA dilutions (lower), coloration of the nitrocellulose membranę with NBT/ BCIP followed by immersion in a toluene bath and fixation onto a glass slide; B) Plot of the absorbance versus optical course for each peak; c: c-Ha-ras fragment; v: v-Ha-ras fragment.

Ando_t K., Saitoh, A., Hino, O., Takahashi, R., Kimura, M. and Katsuki, M., Chemically induced forestomach papillomas in transgenic mice carrying mutant human c-Ha-ras transgenes. Cancer Res. 1992, 52: 978-982.

Barbacid, M., RASgcnes. Ann. Rev. Blochem. 1987,56: 779-827.

Breuer, M._t Wientjens, E., Verbeek, S., Slebos, R. and Berns, A., Carcinogen-induced lymphomagenesis in pim-\ transgenic mice: dose dependence and involvement of myc and ras. Cancer Res. 1991, 51: 958-963.

Dycaico, M.J., Provost, G.S., Kretz, P.L., Ransom, S.L., Moores, J.C. and Short, J.M., The use of shuttle vectors for mutation analysis in transgenic mice and rats. Mutat. Res. 1994,307:461-478.

Ferre, F., Marchese, A., Pezzoli, P., Griffin, S., Buxton, E. and Boyer, V., Quantitative PCR: an overview. In: The Polymerase Chain Reaction. Mullis, K.B., Ferre, F. and Gibbs, R.A. (eds.), Birkhauser, Boston, 1994, pp. 67-88.

Goldsworthy, T.L., Recio, L., Brown, K., Donehower, L.A., Mirsalis, J.C., Tennant, R.W. and Purchase, I.F.H., Symposium overview: transgenic animals in toxicology. Fund. Appl. Toxicol. 1994, 22: 8-19.

Harvey, M., McArthur, M.J., Montgomery, C.A., Butel, J.S., Bradley, A. and Donehower, L.A., Spontaneous and carcinogen-induced tumorigenesis in p53-deficient mice. Naturę Gen. 1993, 5: 225-229.

łzawa, M., Takayama, S., Shindo-Okada, N., Doi, S., Kimura, M., Katsuki, M. and Nishimura, S., Inhibition of Chemical carcinogenesis/n vivo by azatyrosine. Cancer Res. 1992, 52: 1628-1630.

Keith, G., Coulais C., Edorh, A., Bottin, M.C. and Rihn, B., Ethylene glycol monobutyl ether has neither epigenetic nor genotoxic effects in rats and transgenic mice. Occup. Hyg. 1995, accepted: July 25, 1995.

Lanzillo, J.L., Preparation of digoxigenin-labelled probes by the polymerase chain reaction. BioTechniąues 1990, 8: 621-622.

Lawrence, J.B., Singer, R.H. and Marselle, L.M., Highly localized tracks of specific transcripts within interphase nuclei visualized by in situ hybridization. Celi 1989,57: 493-502.

Lech, K. and Brent, R., Resolution and recovery of large DNA fragments. In: Current Protocols in molecular Biology, vol. 1, Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. and Struhl, K. (eds.), Wiley Interscience, New York, 1989, pp. 1.6.5.

Łowy, D.R. and Willumsen, B.M., Function and regulation of RAS. Ann. Rev. Blochem. 1993, 62: 851-891.

Marshall, E., Toxicology goes molecular. Science 1993,259:

116