Abstract
We report the first successful production of transgenic goats that express a heterologous protein in their milk. The production of a glycosylation variant of human tPA (LAtPA - longer acting tissue plasminogen activator) from an expression vector containing the murine whey acid promoter (WAP) operatively linked to the cDNA of a modified version of human tPA was examined in transgenic dairy goats. Two transgenic goats were identified from 29 animals born. The first animal, a female, was mated and allowed to carry the pregnancy to term. Milk was obtained upon parturition and was shown to contain enzymatically active LAtPA at a concentration of 3 μg/ml.
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
Hennighausen, L. 1990. The mammary gland as a bioreactor: production of foreign proteins in milk. Protein Expression and Purification 1: 3–8.
Ebert, K.M. and Selgrath, J.P. 1991. Changes in domestic livestock through genetic engineering. In: Applications of Basic Research in Mammalian Development. Pedersen, R. A., McLaren, A. and First, N. (Eds.). Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.
Gordon, K., Lee, E., Vitale, J.A., Smith, A.E., Westphal, H. and Henninghausen, L. 1987. Production of human tissue plasminogen activator in transgenic mice. Bio/Technology 5: 1183–1187.
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 cDNA to the mammary gland in transgenic mice. Proc. Natl. Acad. Sci. USA 85: 5874–5878.
Clark, A.J., Bessos, H., Bishop, J.O., Brown, P., Harris, S., Lathe, R., McClenaghan, M., Prowse, C., Simons, J.P., Whitelaw, C.B.A. and Wilmut, I. 1989. Expression of human anti-hemophilic factor IX in the milk of transgenic sheep. Bio/Technology 7: 487–492.
Yu, S.H., Deen, K.C., Lee, E., Hennighausen, L., Sweet, R.W., Rosenberg, M. and Westphal, H. 1989. Functional human CD4 protein produced in milk of transgenic mice. Mol. Biol. Med. 6: 255–261.
Buhler, Th.A., Bruyere, Th., Went, D.F., Stranzinger, G. and Burki, K. 1990. Rabbit β-casein promoter directs secretion of human interleukin-2 into the milk of transgenic rabbits. Bio/Technology 8: 140–143.
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.
Meade, H., Gates, L., Lacy, E. and Lonberg, N. 1990. Bovine alpha S1-casein gene sequences direct high level expression of active human urokinase in mouse milk. Bio/Technology 8: 443–446.
Wall, R.J., Pursel, V.G., Shamay, A., McKnight, R.A., Pittius, C.W. and Henninghausen, 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.
Simos, E., Voutsinas, L.P. and Pappas, C.P. 1991. Composition of milk of native Greek goats in the region of Metsovo. Small Ruminant Research 4: 47–60.
Lau, D., Kuzma, G., Wei, C.M., Livingstron, D.J. and Hsiung, N. 1987. A modified human tissue plasminogen activator with extended half-life in vivo. Bio/Technology 5: 953–958.
Selgrath, J.P., Memon, M.A., Smith, T.E. and Ebert, K.M. 1990. Collection and transfer of microinjectable embryos from dairy goats. Theriogenology 34: 1195–1205.
Cuthbertson, R.A. and Klintworth, G.K. 1988. Biology of Disease: Transgenic mice - a gold mine for furthering knowledge in pathobiology. Laboratory Investigation 58: 484–502.
Ebert, K.M., Smith, T.E., Buonomo, F.C., Overstrom, E.W. and Low, M.J. 1990. Porcine growth hormone gene expression from viral promoters in transgenic swine. Animal Biotechnology 1: 145–159.
Hammer, R.E., Pursel, V.G., Rexroad, C.E., Wall, R.J., Bolt, D.J., Ebert, K.M., Palmiter, R.D. and Brinster, R.L. 1985. Production of transgenic rabbits, sheep and pigs by microinjection. Nature 315: 680–683.
Feinberg, A. and Vogelstein, B. 1983. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 72: 248–254.
Thomas, P. 1980. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc. Natl. Acad. Sci. 77: 5201–5205.
Bradford, M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248–254.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ebert, K., Selgrath, J., DiTullio, P. et al. Transgenic Production of a Variant of Human Tissue-Type Plasminogen Activator in Goat Milk: Generation of Transgenic Goats and Analysis of Expression. Nat Biotechnol 9, 835–838 (1991). https://doi.org/10.1038/nbt0991-835
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/nbt0991-835
This article is cited by
-
Generation of goats by nuclear transfer: a retrospective analysis of a commercial operation (1998–2010)
Transgenic Research (2020)
-
Intratesticular injection followed by electroporation allows gene transfer in caprine spermatogenic cells
Scientific Reports (2018)
-
Genome editing and genetic engineering in livestock for advancing agricultural and biomedical applications
Mammalian Genome (2017)
-
High-level expression of a novel recombinant human plasminogen activator (rhPA) in the milk of transgenic rabbits and its thrombolytic bioactivity in vitro
Molecular Biology Reports (2016)
-
Ingestion of milk containing the Dp2 peptide, a dust mite allergen, protects mice from allergic airway inflammation and hyper-responsiveness
Allergy, Asthma & Clinical Immunology (2013)
