Massoud M, Attal J, Thépot D, Pointu H, Stinnakre M G, Théron M C, Lopez C, Houdebine L M
Unité d'endocrinologie de l'embryon, Inra, Jouy-en-Josas, France.
Reprod Nutr Dev. 1996;36(5):555-63. doi: 10.1051/rnd:19960511.
Human erythropoietin (EPO) gene and cDNA associated with the rabbit whey acidic protein (WAP) gene promoter were used to tentatively produce the recombinant protein in milk of transgenic mice and rabbits. Several gene constructs showed good efficiency in the mouse mammary cell line HC11. None of them was able to direct the expression of the hormone at a concentration higher than 50 micrograms/mL in mouse and rabbit milk. With one of the construct, the rabbits had an abnormally high amount of red blood cells irrespectively of their sex, they could not reproduce and no milk could be obtained from them. These animals died prematurely. In these animals, the EPO gene was therefore expressed at a low but supraphysiological level in organs other than the mammary gland. These experiments show that transgenic animals obtained with gene constructs which do not contain insulators cannot be used as living fermentors to produce human erythropoietin in their milk at an industrial scale.
将与兔乳清酸性蛋白（WAP）基因启动子相关的人促红细胞生成素（EPO）基因和cDNA用于尝试在转基因小鼠和兔的乳汁中生产重组蛋白。几种基因构建体在小鼠乳腺细胞系HC11中显示出良好的效率。但没有一种能够在小鼠和兔乳中指导激素表达的浓度高于50微克/毫升。使用其中一种构建体时，兔子无论性别，红细胞数量都异常高，它们无法繁殖，也无法从它们身上获取乳汁。这些动物过早死亡。因此，在这些动物中，EPO基因在乳腺以外的器官中以低但超生理水平表达。这些实验表明，用不含绝缘子的基因构建体获得的转基因动物不能用作活体发酵罐以工业规模在其乳汁中生产人促红细胞生成素。
