Transgenic rabbits carrying gene constructs encoding human nerve growth factor beta (hNGF-beta) cDNA were generated. Expression of hNGF-beta mRNA was restricted to the mammary gland of lactating rabbits. Western Blot analysis revealed a polypeptide of 13.2 kDa in the milk of transgenic animals, hNGF-beta was purified from the milk by a two-step chromatographic procedure, Electrospray mass spectroscopy analysis of purified hNGF-beta depicted a molecular weight of 13 261 Da per subunit. The biological activity of the hNGF-beta was tested using PC12W2 cells and cultures of dorsal root ganglion neurons from chicken embryos. Crude defatted milk from transgenic animals and purified hNGF-beta demonstrated full biological activity when compared to commercial recombinant hNGF-beta. (C) 1999 Federation of European Biochemical Societies.
Animals - : Animals, Genetically Modified - : Cell Size - drug effects : Cells, Cultured - drug effects : Chick Embryo - drug effects : Dose-Response Relationship, Drug - drug effects : Female - drug effects : Ganglia, Spinal - cytology : Humans - cytology : Lactation - metabolism : Male - metabolism : Mammary Glands, Animal - metabolism : Mass Spectrometry - metabolism : Milk - metabolism : Nerve Growth Factors - chemistry : Neurites - drug effects : Organ Specificity - drug effects : PC12 Cells - drug effects : RNA, Messenger - analysis : Rabbits - analysis : Rats - analysis : Recombinant Proteins - chemistry