Production of biologically active human granulocyte colony stimulating factor in the milk of transgenic goat.
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.
Ko
J H
JH
Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Taejon.
Lee
C S
CS
Kim
K H
KH
Pang
M G
MG
Koo
J S
JS
Fang
N
N
Koo
D B
DB
Oh
K B
KB
Youn
W S
WS
Zheng
G D
GD
Park
J S
JS
Kim
S J
SJ
Han
Y M
YM
Choi
I Y
IY
Lim
J
J
Shin
S T
ST
Jin
S W
SW
Lee
K K
KK
Yoo
O J
OJ
D000818:Animals; D030801:Animals, Genetically Modified; D015153:Blotting, Western; D002364:Caseins / Q000235:genetics; D002455:Cell Division; D003520:Cyclophosphamide / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005822:Genetic Vectors; D006041:Goats / Q000502:physiology*; D016179:Granulocyte Colony-Stimulating Factor / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008892:Milk / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
D002364:Caseins; D017931:DNA Primers; D011993:Recombinant Fusion Proteins; D016179:Granulocyte Colony-Stimulating Factor; D003520:Cyclophosphamide