人促血小板生成素在转基因小鼠乳腺中定位表达的研究(英文)

通过转基因动物乳腺生物反应器大规模生产药用蛋白质已成为现代生物技术新的生长点之一。为研制表达人促血小板生成素的哺乳动物生物反应器的转基因小鼠模型，本论文以小鼠乳清酸蛋白 (mWAP) 基因5挾说骺厍团-s1-酪蛋白基因3挾说骺厍魑鹘谠菇擞糜诒泶锶舜傺“迳伤氐娜橄僮橹匾煨员泶镌靥錺WAPTPO(Fig.1)。通过常规显微注射的方法把mWAP启动子指导的hTPO表达载体导入小鼠受精卵，获得出生小鼠16只。经PCR检测，有6只为转基因阳性(Fig.2)。G0代小鼠中转基因整合率为37.5% (6/16)，用ELISA方法在G0代转基因雌鼠的乳汁中检测了促血小板生成素的表达，表达量在0.8 mg/mL以上(Table 1)。这些结果表明我们已建立了乳腺表达hTPO 的转基因小鼠模型，为以后大型家畜乳腺生物反应器的研制提供了科学依据。

Expression of Human Thrombopoietin in Milk of Transgenic Mice

Producing of pharmaceutical proteins in milk of transgenic animals is one of the most promising areas in modern biotechnology. To establish transgenic mice model of mammary gland bioreactor for production of human recombinant thrombopoietin, the mammary gland-specific expression vector pWAPTPO was constructed using 5?regulatory sequence of whey acidic protein (mWAP) gene and 3?control region of bovine a-S1-casein gene. The chimeric gene was injected into mouse zygotes by regular pronuclear microinjection. 16 G0 generation founder mice were obtained. The integration analysis indicates that 6 of them were transgenic, thus the rate of integration was 37.5% (6/16). The presence of recombinant human thrombopoietin was detected in the milk of transgenic mice at levels of up to 0.8 mg/mL. These results showed that the established transgenic mouse model could produce recombinant proteins in the milk of lactating females.