扬州大学教师个人主页服务平台袁玉国--中文主页-- High-level expression of a novel recombinant human plasminogen activator (rhPA) in the milk of transgenic rabbits and its thrombolytic bioactivity in vitro

袁玉国

学位：博士
职称：副高级
所在单位：兽医学院

博士生导师

硕士生导师

教师英文名称：Yuguo Yuan
教师拼音名称：Yuan Yuguo
电子邮箱：
入职时间：2010-06-01
学历：博士研究生毕业
办公地点：扬州大学文汇路校区45楼102室
性别：男
联系方式：0514-87979228
在职信息：在岗
毕业院校：扬州大学

学科：临床兽医学

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High-level expression of a novel recombinant human plasminogen activator (rhPA) in the milk of transgenic rabbits and its thrombolytic bioactivity in vitro

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发表刊物：

Mol Biol Rep

摘要：

The human tissue-type plasminogen activator (tPA) is a key kinase of fibrinolysis that plays an important role in dissolving fibrin clots to promote thrombolysis. The recombinant human plasminogen activator (rhPA) has more thrombolytic advantages than the wild type tPA. To increase the half-life and thrombolytic activity of tPA, a mutant containing only the essential K2 fibrin-binding and P activating plasminogen domains of the wild type tPA was cloned. This fragment was then inserted into goat β-casein regulatory sequences. Then, a mammary gland-specific expression vector, PCL25/rhPA, was constructed, and the transgenic rabbits were generated. In this study, 18 live transgenic founders (12♀, 6♂) were generated using pronuclear microinjection. Six transgenic rabbits were obtained, and the expression levels of rhPA in the milk had a range of 15.2-630 µg/ml. A fibrin agarose plate assay of rhPA showed that it had strong thrombolytic bioactivity in vitro, and the highest specific activity was >360 (360 times more than that of alteplase). The results indicated that the rhPA containing only the K2 and P domains is efficiently expressed with higher thrombolytic bioactivity in the milk of transgenic rabbits. Our study also demonstrated a new method for the large-scale production of clinically relevant recombinant pharmaceutical proteins in the mammary glands of transgenic rabbits.

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否

发表时间：

2016-08-23

收录刊物：

SCI

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个人简介

袁玉国，博士，副教授，硕士、博士生导师。2010年毕业于扬州大学兽医学院，获临床兽医学博士学位，并留校任教。2015.10—2017.7韩国Konkuk University博士后，2017.9—2018.9韩国Gyeongsang National University访问学者。

长期从事动物体细胞克隆和转基因克隆的研究工作，2007年在国内外首次报道利用成年乳腺上皮细胞获得克隆山羊，先后获得转人乳铁蛋白，白蛋白等转基因克隆羊。利用TALEN，Cas9技术获得Fel d1基因敲除猫、MSTN基因敲除山羊，人高血脂兔动物模型（兔）等。同时开展了功能纳米材料的制备及其在抗菌、抗肿瘤和化妆品上的应用。先后主持了国家自然科学基金、国家转基因重大专项子课题、江苏省种业“揭榜挂帅”等15余项课题的研究工作，在 EBioMedicine，Int J Nanomedicine，Cells，Oxid Med Cell Longev, Int J Mol Sci，Mol Reprod Dev，Theriogenology等杂志发表SCI论文30余篇，获得省部级等科技奖励2项、发明专利1项。

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