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| 蚓激酶重组逆转录病毒载体pLN-BCP-LK~R的构建及其在山羊乳腺组织中的表达 | |
| 引用本文： | 姚伟,任文陟,张守峰,范志强,扈荣良,涂长春.蚓激酶重组逆转录病毒载体pLN-BCP-LK~R的构建及其在山羊乳腺组织中的表达[J].中国生化药物杂志,2005,26(3):129-132. |
| 作者姓名： | 姚伟 任文陟 张守峰 范志强 扈荣良 涂长春 |
| 作者单位： | 1. 军事医学科学院,军事兽医研究所;吉林大学,实验动物中心,吉林,长春,130062 2. 吉林大学,实验动物中心,吉林,长春,130062 3. 军事医学科学院,军事兽医研究所 |
| 基金项目： | 国家"863"高技术研究发展计划(No.2002AA206653)基金资助 |
| 摘 要： | 目的在山羊奶中表达蚓激酶。方法将含有蚓激酶基因的表达盒插入到逆转录病毒载体pLNCL中获得了蚓激酶重组逆转录病毒载体,转染泌乳期奶山羊乳腺小叶并获得暂时表达。利用脂质体转染PA317包装细胞,G418进行筛选得到阳性细胞克隆,克隆扩大培养后病毒上清感染妊娠期奶山羊乳腺小叶组织,山羊产后采奶纤维蛋白平板溶圈法检测蚓激酶活性。结果产后奶样经纤维蛋白平板溶圈法检测羊奶具有明显纤溶活性。结论蚓激酶基因已经整合到奶山羊乳腺组织并能实现相对稳定的表达。 |
| 关 键 词： | 蚓激酶 重组逆转录病毒 包装 转染 表达 |
| 文章编号： | 1005-1678(2005)03-0129-04 |
| 修稿时间： | 2004年9月29日 |
Construction of recombinant retroviral vector containing lumbrokinase gene and its expression in goat gland |
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| YAO Wei,REN Wen-zhi,ZHANG Shou-feng,FAN Zhi-qiang,HU Rong-liang,TU Chang-Chun.Construction of recombinant retroviral vector containing lumbrokinase gene and its expression in goat gland[J].Chinese Journal of Biochemical Pharmaceutics,2005,26(3):129-132. | |
| Authors: | YAO Wei REN Wen-zhi ZHANG Shou-feng FAN Zhi-qiang HU Rong-liang TU Chang-Chun |
| Institution: | YAO Wei,REN Wen-zhi,ZHANG Shou-feng,FAN Zhi-qiang,HU Rong-liang,TU Chang-chun Military Veterinary Institute,Academy of Military Medical Science,Laboratory Animal Center,Jilin University,Changchun 130062,China |
| Abstract: | Purpose To express lumbrokinase(LK) in goat gland. Methods LK gene was cloned into the Retroviral Vector pLNCL to construct the recombinant retroviral vector pLN-BCP-LKR. The result of fibrin agrose plate assay(FAPA) showed that LK gene was expressed in the milk by injecting the plasmid pLN-BCPLK into the galactophore of female goat. The vector was transfected into PA317 cells for packaging by liposomal transfection. After G418 selection for 2 weeks,G418-resistant PA317 colonies were obtained and amplified. The supernatant of cell culture was harvested and injected into the mammary gland of pregnant goat. Results The milk displayed relatively high fibrinolysis activity in vitro on FAPA. Conclusion LK gene had been integrated into the mammary gland and expressed continuously in the milk of the post partum goat. |
| Keywords: | lumbrokinase recombinant retrovirus package transfection expression |
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