Ko J H, Lee C S, Kim K H, Pang M G, Koo J S, Fang N, Koo D B, Oh K B, Youn W S, Zheng G D, Park J S, Kim S J, Han Y M, Choi I Y, Lim J, Shin S T, Jin S W, Lee K K, Yoo O J
Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Taejon.
Transgenic Res. 2000 Jun;9(3):215-22. doi: 10.1023/a:1008972010351.
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.
我们通过将山羊β-酪蛋白启动子/人粒细胞集落刺激因子(G-CSF)融合基因显微注射到单细胞山羊受精卵中,培育出了一只转基因雌性山羊。转基因山羊奶中产生的人G-CSF水平高达50微克/毫升。在使用体外HL-60细胞增殖进行检测时,其生物学活性与中国仓鼠卵巢(CHO)细胞表达的重组人G-CSF相当。转基因山羊奶中的人G-CSF增加了环磷酰胺(CPA)诱导的白细胞减少的C57BL/6N小鼠的白细胞总数。分泌的人G-CSF进行了糖基化修饰,尽管与CHO细胞来源的人G-CSF相比,O-糖基化程度较低。