Human nerve growth factor beta (hNGF-beta): mammary gland specific expression and production in transgenic rabbits

FEBS Lett. 1999 Feb 5;444(1):111-6. doi: 10.1016/s0014-5793(98)01728-1.

Abstract

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.

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Cell Size / drug effects
  • Cells, Cultured
  • Chick Embryo
  • Dose-Response Relationship, Drug
  • Female
  • Ganglia, Spinal / cytology
  • Humans
  • Lactation / metabolism
  • Male
  • Mammary Glands, Animal / metabolism*
  • Mass Spectrometry
  • Milk / metabolism
  • Nerve Growth Factors / chemistry
  • Nerve Growth Factors / genetics
  • Nerve Growth Factors / metabolism*
  • Nerve Growth Factors / pharmacology
  • Neurites / drug effects
  • Neurites / metabolism
  • Organ Specificity
  • PC12 Cells
  • RNA, Messenger / analysis
  • Rabbits
  • Rats
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / pharmacology

Substances

  • Nerve Growth Factors
  • RNA, Messenger
  • Recombinant Proteins