Abstract
We have produced a line of transgenic mice carrying a hybrid bovine αs1 casein/human urokinase gene. Bovine αs1-casein gene regulatory sequences specifically direct expression of the human urokinase gene in lactating mammary tissue from these mice. Urokinase is a 54 kD protein with 9 disulfide bonds that is normally synthesized in the kidney; however, the casein/urokinase transgenic mice secrete active human urokinase into their milk at concentrations of 1-2 mg/ml. The mice show no other abnormalities. They give birth to, and nurse, normal sized healthy litters. Thus it is possible to produce high concentrations of a large, cysteine rich, non-milk protein in the milk of transgenic animals. This line of transgenic mice provides a model for the eventual production of transgenic farm animals producing high levels of recombinant proteins in their milk.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on SpringerLink
- Instant access to the full article PDF.
USD 39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Simons, J.P., McClenaghan, M. and Clark, A.J. 1987. Alteration of the quality of milk by expression of sheep β-lacoglobulin in transgenic mice. Nature 328: 530–532.
Andres, A.C., Schoenenberger, C.A., Groner, B., Hennighausen, L., Lemeur, M. and Gerlinger, P. 1987. Ha-ras oncogene expression directed by a milk protein gene promoter. Proc. Natl. Acad. Sci. U.S.A. 84: 1299–1303.
Pittius, C.W., Henninghausen, L., Lee, E., Westphal, H., Nicols, E., Vitale, J. and Gordon, K. 1988. A milk protein gene promoter directs the expression of human tissue plasminogen activator complementary DNA to the mammary gland in transgenic mice. Proc. Natl. Acad. Sci. U.S.A. 85: 5874–5878.
Gordon, K., Lee, E., Vitale, J., Smith, A.E., Westphal, H. and Henninghausen, L. 1987. Production of human tissue plasminogen activator in transgenic mice. Bio/Technology 5: 11831–1185.
Willis, I.M., Stewart, A.F., Caputo, A., Thompson, A.R. and Mac-Kinlay, A.G. 1982. Construction and identification by partial nucleotide sequence analysis of bovine casein and β-lactoglobulin cDNA clones. DNA 1: 375–386.
Jenness, R., 1970. Milk Proteins,Chemistry and Molecular Biology, p. 25, Vol. I, H. A. McKenzie (Ed.). Academic Press, New York
Hogan, B., Costantini, F. and Lacy, E. 1986. Methods of Manipulating the Mouse Embryo. Cold Spring Harbor Laboratory, New York.
Verde, P., Stoppelli, M.P., Galeffi, P., Di Nocera, P. and Blasi, F. 1984. Identification and primary sequence of an unspliced human uroki-lase poly(A)+ RNA. Proc. Natl. Acad. Sci. U.S.A. 81: 4727–4731.
Nagamine, Y., Sudol, M. and Reich, E. 1983. Hormonal regulation of plasminogen activator mRNA production in porcine kidney cells. Cell 32: 1181–1190.
Hohn, B. and Collins, J. 1980. A small cosmid for efficien cloning of large DNA fragments. Gene 11: 291–298.
Grosveld, F.G., Lund, T., Murray, E.J., Mellor, A.L., Dahl, H. and Flavell, R. 1982. The construction of cosmid libraries which can be used to transform eukaryotic cells. Nucleic Acids Res. 10: 6715–6732.
Riccio, A., Grimaldi, G., Verde, P., Sebastio, G., Boast, S., Blasi, F., 1985. The human urokinase-plasminogen activator gene and its promoter. Nucleic Acids Res. 13: 2759–2770.
Chomczynski, P. and Sacchi, N. 1987. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Analytical Biochem. 162: 156–159.
Church, G. and Gilbert, W. 1984. Genomic sequencing. Proc. Natl. Acad. Sci. U.S.A. 81: 1991–1995.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Meade, H., Gates, L., Lacy, E. et al. Bovine AlphaS1-Casein Gene Sequences Direct High Level Expression of Active Human Urokinase in Mouse Milk. Nat Biotechnol 8, 443–446 (1990). https://doi.org/10.1038/nbt0590-443
Received:
Accepted:
Issue date:
DOI: https://doi.org/10.1038/nbt0590-443
This article is cited by
-
A 3,387 bp 5′-flanking sequence of the goat alpha-S1-casein gene provides correct tissue-specific expression of human granulocyte colony-stimulating factor (hG-CSF) in the mammary gland of transgenic mice
Transgenic Research (2012)
-
Regulation of human protein C gene expression by the mouse WAP promoter
Transgenic Research (1994)
-
Expression of human lactoferrin in milk of transgenic mice
Transgenic Research (1994)
-
Expression of human serum albumin in the milk of transgenic mice
Transgenic Research (1992)
