Human nerve growth factor beta (hNGF-beta): mammary gland specific expression and production in transgenic rabbits.
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.
Ludwig Boltzmann Institute for Immuno-, Cyto- and Molecular Genetic Research, Vienna, Austria.
S
S
Coulibaly
U
U
Besenfelder
M
M
Fleischmann
N
N
Zinovieva
A
A
Grossmann
M
M
Wozny
I
I
Bartke
M
M
Tögel
M
M
Müller
G
G
Brem
D000818:Animals; D030801:Animals, Genetically Modified; D048429:Cell Size / Q000187:drug effects; D002478:Cells, Cultured; D002642:Chick Embryo; D004305:Dose-Response Relationship, Drug; D005260:Female; D005727:Ganglia, Spinal / Q000166:cytology; D006801:Humans; D007774:Lactation / Q000378:metabolism; D008297:Male; D008321:Mammary Glands, Animal / Q000378:metabolism*; D013058:Mass Spectrometry; D008892:Milk / Q000378:metabolism; D009414:Nerve Growth Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D016501:Neurites / Q000187:drug effects / Q000378:metabolism; D009928:Organ Specificity; D016716:PC12 Cells; D012333:RNA, Messenger / Q000032:analysis; D011817:Rabbits; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology
D009414:Nerve Growth Factors; D012333:RNA, Messenger; D011994:Recombinant Proteins