重组腺病毒介导的人尿激酶原突变体在山羊乳汁中的表达

构建携带人尿激酶原突变体cDNA的重组腺病毒，通过该腺病毒介导实现外源基因在山羊乳腺中表达。通过大肠杆菌内同源重组将人尿激酶原突变体cDNA插入到腺病毒载体中，经过293细胞包装获得重组腺病毒，直接注射到泌乳山羊乳腺，收集感染后1~4天的乳汁，利用纤维蛋白溶圈试验、Western blot、ELISA检测乳清中尿激酶原突变体的表达。结果显示病毒注射后1~4天乳清中均可检测到尿激酶原的表达，其表达量可达0.41mg/ml。该方法可以实现重组蛋白在山羊乳腺的短期表达，可能是大规模生产临床医疗蛋白的一条经济有效的途径。

Expression of Human Prourokinase Mutant in the Milk of Goat Mediated by the Recombinant Adenovirus

In order to express the recombinant proteins in the milk of goat, a recombinant adenovirus was constructed in which the Pro-urokinase mutant cDNA was cloned into the adenovirus vector, and then was transfected to 293 for packaging. The recombinant adenovirus were directly injected into the goat mammary glands, the milk serum was collected 1-4d post-infection and then subjected to fibrin plate assay，Western blotting and ELISA assay. The results showed that the human pro-urokinase was expressed efficiently in the milk of goat, even attained to 0.41mg/mL in the 2rd day post-infection. Taken together, the method is quite useful for the transient expression of recombinant proteins, and the approach established here is probably one of the most economical and efficient ways for large-scale production of recombinant proteins of biopharmaceutical interest.