Production of biologically active human granulocyte colony stimulating factor in the milk of transgenic goat

Transgenic Res. 2000 Jun;9(3):215-22. doi: 10.1023/a:1008972010351.

Abstract

We have developed a transgenic female goat harboring goat beta-casein promoter/human granulocyte colony stimulating factor (G-CSF) fusion gene by microinjection into fertilized one-cell goat zygotes. Human G-CSF was produced at levels of up to 50 microg/ml in transgenic goat milk. Its biological activity was equivalent to recombinant human G-CSF expressed from Chinese hamster ovary (CHO) cell when assayed using in vitro HL-60 cell proliferation. Human G-CSF from transgenic goat milk increased the total number of white blood cells in C57BL/6N mice with leucopenia induced by cyclophosphamide (CPA). The secreted human G-CSF was glycosylated although the degree of O-glycosylation was lower compared to CHO cell-derived human G-CSF.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified*
  • Blotting, Western
  • Caseins / genetics
  • Cell Division
  • Cyclophosphamide / metabolism
  • DNA Primers / chemistry
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Genetic Vectors
  • Goats / physiology*
  • Granulocyte Colony-Stimulating Factor / biosynthesis*
  • Granulocyte Colony-Stimulating Factor / genetics
  • Granulocyte Colony-Stimulating Factor / isolation & purification
  • Humans
  • Mice
  • Mice, Inbred C57BL
  • Milk / metabolism*
  • Polymerase Chain Reaction
  • Recombinant Fusion Proteins / biosynthesis*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / isolation & purification

Substances

  • Caseins
  • DNA Primers
  • Recombinant Fusion Proteins
  • Granulocyte Colony-Stimulating Factor
  • Cyclophosphamide