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Human nerve growth factor beta (hNGF‐β): mammary gland specific expression and production in transgenic rabbits
Author(s) -
Coulibaly S,
Besenfelder U,
Fleischmann M,
Zinovieva N,
Grossmann A,
Wozny M,
Bartke I,
Tögel M,
Müller M,
Brem G
Publication year - 1999
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)01728-1
Subject(s) - nerve growth factor , complementary dna , microbiology and biotechnology , biology , mammary gland , transgene , dorsal root ganglion , recombinant dna , western blot , embryo , genetically modified mouse , northern blot , endocrinology , medicine , gene , dorsum , anatomy , biochemistry , receptor , genetics , cancer , breast cancer
Transgenic rabbits carrying gene constructs encoding human nerve growth factor beta (hNGF‐β) cDNA were generated. Expression of hNGF‐β 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‐β was purified from the milk by a two‐step chromatographic procedure. Electrospray mass spectroscopy analysis of purified hNGF‐β depicted a molecular weight of 13 261 Da per subunit. The biological activity of the hNGF‐β was tested using PC12W2 cells and cultures of dorsal root ganglion neurons from chicken embryos. Crude defatted milk from transgenic animals and purified hNGF‐β demonstrated full biological activity when compared to commercial recombinant hNGF‐β.