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
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Animals, Genetically Modified*
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Blotting, Western
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Caseins / genetics
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Cell Division
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Cyclophosphamide / metabolism
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DNA Primers / chemistry
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Enzyme-Linked Immunosorbent Assay
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Female
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Genetic Vectors
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Goats / physiology*
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Granulocyte Colony-Stimulating Factor / biosynthesis*
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Granulocyte Colony-Stimulating Factor / genetics
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Granulocyte Colony-Stimulating Factor / isolation & purification
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Humans
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Mice
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Mice, Inbred C57BL
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Milk / metabolism*
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Polymerase Chain Reaction
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Recombinant Fusion Proteins / biosynthesis*
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / isolation & purification
Substances
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Caseins
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DNA Primers
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Recombinant Fusion Proteins
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Granulocyte Colony-Stimulating Factor
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Cyclophosphamide