Bio/Technology
9, 835 - 838 (1991)
doi:10.1038/nbt0991-835
Transgenic Production of a Variant of Human Tissue-Type Plasminogen Activator in Goat Milk: Generation of Transgenic Goats and Analysis of ExpressionKarl M. Ebert1, *, James P. Selgrath1, Paul DiTullio2, Julie Denman2, Thomas E. Smith1, Mushtaq A. Memon1, Joanne E. Schindler1, Glen M. Monastersky2, James A. Vitale2
& Katherine Gordon2, 3
1Tufts University, Schools of Veterinary Medicine, Medicine, and Dental Medicine, 200 Westboro Road, North Grafton, MA 01536-1895.
2Genzyme Corporation, One Mountain Road, Framingham, MA 01701.
3Current address: Biodynamics, 222 Third Street, Suite 3121, Cambridge, MA 02142.
*Corresponding author. 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 g/ml. REFERENCES
- Hennighausen, L. 1990. The mammary gland as a bioreactor: production of foreign proteins in milk. Protein Expression and Purification 1: 3−8. | PubMed | ChemPort |
- 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. | ISI | ChemPort |
- 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. | PubMed | ChemPort |
- 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. | ISI | ChemPort |
- 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. | PubMed | ISI | ChemPort |
- Buhler, Th.A., Bruyere, Th., Went, D.F., Stranzinger, G. and Burki, K. 1990. Rabbit
-casein promoter directs secretion of human interleukin-2 into the milk of transgenic rabbits. Bio/Technology 8: 140−143. | PubMed | ISI | ChemPort |
- Archibald, A.L., McClenaghan, M., Hornsey, V., Simons, J.P. and Clark, A.J. 1990. High-level expression of biologically active human
1-antitrypsin in the milk of transgenic mice. Proc. Natl. Acad. Sci. USA 87: 5178−5182. | PubMed | ChemPort |
- 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. | PubMed | ISI | ChemPort |
- 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. | PubMed | ChemPort |
- 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. | Article | ISI |
- 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. | ISI | ChemPort |
- 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. | ISI |
- 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. | PubMed | ISI | ChemPort |
- 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. | ChemPort |
- 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. | PubMed | ISI | ChemPort |
- 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. | PubMed | ChemPort |
- 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. | Article | PubMed | ISI | ChemPort |
|