Abstract
We describe the generation of five sheep transgenic for a fusion of the ovine β-lactoglobulin gene promoter to the human α1-antitrypsin (hα1AT) genomic sequences. Four of these animals are female and one male. Analysis of the expression of hα1AT in the milk of three of these females shows that all express the human protein at levels greater than 1 gram per liter. In one case initial levels exceeded 60 grams per liter and stabilized at approximately 35 grams per liter as lactation progressed. Human α1AT purified from the milk of these animals appears to be fully N-glycosylated and has a biological activity indistinguishable from human plasma-derived material.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Palmiter, R.D., Brinster, R.L., Hammer, R.E., Trumbauer, M.E., Rosenfeld, M.G., Birnberg, N.C. and Evans, R.M. 1982. Dramatic growth of mice that develop from eggs microinjected with metallothionein-growth hormone fusion genes. Nature 300: 611–615.
Lovell-Badge, R.H. 1985. New advances in the field. Nature 315: 628–629.
Hennighausen, L., Ruiz, L. and Wall, R. 1990. Transgenic animals-production of foreign proteins in milk. Curr. Op. in Biotech. 1: 74–78.
Simons, J.P., McClenaghan, M. and Clark, A.J. 1987. Alteration of the quality of milk by expression of sheep β-lactoglobulin in transgenic mice. Nature 328: 530–532.
Wall, R.J., Pursel, V.G., Shamay, A., McKnight, R.A., Pittius, C.W. and Hennighausen, L. 1991. High-level synthesis of a heterologous milk protein in the mammary glands of transgenic swine. Proc. Natl. Acad. Sci. USA 88: 1696–1700.
Meade, H., Gates, L., Lacy, E. and Lonberg, N. 1990. Bovine αS1-casein gene sequences direct high level expression of active human urokinase in mouse milk. Bio/Technology 8: 443–446.
Crystal, R.G. 1989. The α1-antitrypsin gene and its deficiency states. Trends in Genet. 5: 411–417.
Carlson, J.A., Rogers, B.B., Sifers, R.N., Hawkins, H.K., Finegold, M.J. and Woo, S.L.C. 1988. Multiple tissues express α1-antitrypsin in transgenic mice and man. J. Clin. Invest. 82: 26–36.
Kelsey, G.D., Povey, S., Bygrave, A.E. and Lovell-Badge, R.H. 1987. Species- and tissue-specific expression of human α1-antitrypsin in transgenic mice. Genes and Development 1: 161–171.
Casolaro, M.A., Fells, G., Wewers, M., Pierce, J.E., Ogushi, F., Hubbard, R., Sellers, S., Forstrom, J., Lyons, D., Kawasaki, G. and Crystal, R.G. 1987. Augmentation of lung antineutrophil elastase capacity with recombinant human α1-antitrypsin. J. Appl. Physiol. 63: 2015–2023.
Archibald, A.L., McClenaghan, M., Hornsey, V., Simons, J.P. and Clark, A.J. 1990. High-level expression of biologically active human α1-antitrypsin in the milk of transgenic mice. Proc. Natl. Acad. Sci. USA 87: 5178–5182.
Simons, J.P., Wilmut, I., Clark, A.J., Archibald, A.L., Bishop, J.O. and Lathe, R. 1988. Gene transfer into sheep. Bio/Technology 6: 179–183.
Grosveld, F., van Assendelft, G.B., Greaves, D.R. and Kollias, G. 1987. Position-independent, high-level expression of the human β-globin gene in transgenic mice. Cell 51: 975–985.
Greaves, D.R., Wilson, F.D., Lang, G. and Kioussis, D. 1989. Human CD2 3′-flanking sequences confer high-level, T cell-specific, position-independant gene expression in transgenic mice. Cell 56: 979–986.
Bonifer, C., Vidal, M., Grosveld, F. and Sippel, A. 1990. Tissue specific and position independent expression of the complete gene domain for chicken lysozyme in transgenic. mice. EMBO J. 9: 2843–2848.
Bishop, J.O. and Al-Shawi, R. 1989. Gene expression in transgenic animals, p. 249–260. In: Evolution and Animal Breeding. Hill, W. G. and Mackay, T. F. C., (Eds.). CAB International, Wallingford, UK.
Brinster, R.L., Alien, J.M., Behringer, R.R., Gelinas, R.E. and Palmiter, R.D. 1988. Introns increase transcriptional efficiency in transgenic mice. Proc. Natl. Acad. Sci. USA 85: 836–840.
Palmiter, R.D., Sandgren, E.P., Avarbock, M.R., Alien, D.D. and Brinster, R.L. 1991. Heterologous introns can enhance expression of transgenes in mice. Proc. Natl. Acad. Sci. USA 88: 478–482.
Choi, T., Huang, M., Gorman, C. and Jaenisch, R. 1991. A generic intron increases gene expression in transgenic mice. Mol. Cell. Biol. 11: 3070–3074.
Courtney, M., Buchwalder, A., Tessier, L-H., Jaye, M., Benavente, A., Balland, A., Kohli, V., Lathe, R., Tolstoshev, P. and Lecocq, J.-P. 1984. High-level production of biologically active human α1-antitrypsin in Escherichia coli. Proc. Natl. Acad. Sci. USA 81: 669–673.
Garver, R.I., Chytil, A., Karlsson, S., Fells, G.A., Brantly, M.L., Courtney, M., Kantoff, P.W., Nienhuis, A.W., French Anderson, W. and Crystal, R.G. 1987. Production of glycosylated physiologically “normal” human α1-antitrypsin by mouse fibroblasts modified by insertion of a human α1-antitrypsin cDNA using a retroviral vector. Proc. Natl. Acad. Sci. USA 84: 1050–1054.
Sleep, D., Belfield, G.P., Ballance, D.J., Steven, J., Jones, S., Evans, L.R., Moir, P.D. and Goody, A.R. 1991. Saccharomyces cerevisiae strains that overexpress heterologous proteins. Bio/Technology 9: 183–187.
Pavironi, A., Skern, T., Le Meur, M., Lutz, Y., Lathe, R., Crystal, R.G., Fuchs, J.-P., Gerlinger, P. and Courtney, M. 1989. Recombinant proteins of therapeutic interest expressed by lymphoid cell lines derived from transgenic mice. Bio/Technology 7: 1049–1054.
Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 277: 680–685.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Wright, G., Carver, A., Cottom, D. et al. High Level Expression of Active Human Alpha-1-Antitrypsin in the Milk of Transgenic Sheep. Nat Biotechnol 9, 830–834 (1991). https://doi.org/10.1038/nbt0991-830
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/nbt0991-830
This article is cited by
-
Generation of goats by nuclear transfer: a retrospective analysis of a commercial operation (1998–2010)
Transgenic Research (2020)
-
Construction of a recombinant human insulin expression vector for mammary gland-specific expression in buffalo (Bubalus bubalis) mammary epithelial cell line
Molecular Biology Reports (2014)
-
Production of transgenic goats expressing human coagulation factor IX in the mammary glands after nuclear transfer using transfected fetal fibroblast cells
Transgenic Research (2013)
-
Alpha-1 Antitrypsin Deficiency: New Developments in Augmentation and Other Therapies
BioDrugs (2013)
-
Could protein tertiary structure influence mammary transgene expression more than tissue specific codon usage?
Transgenic Research (2010)