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人组织型纤溶酶原激活剂乳腺定位表达载体的构建及其表达 被引量:3

Construction of Mammary specific Expressing Vectors of Human Tissue type Plasminogen Activator and Expression in Milk of Rabbit
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摘要 将编码人组织型纤溶酶原激活剂(tPA) 的1766bp 基因片段插入大鼠β-乳酪蛋白 (β-Casein) 启动子的下游及pSVL质粒的PolyA上游。构建了pβ-Casein-PA1、pβ-Casein-PA2 两个乳腺定位表达载体。将其分别与脂质体混合制备转染液后, 直接注入妊娠29 天的青紫蓝母兔乳腺中。结果表明:两种载体均能在乳腺中表达,且具有溶纤活性,表达量在200—500ng/m l之间。这不仅证明了所构建载体可用于转基因乳腺生物反应器的建立, 而且提示, The human tissue plasminogen activator (tPA) cDNA with the length of 1766bp gene which encodes the complete tPA was inserted into the downstream of rat beta casein and upstream of the poly(A) of pSVL plasmid to construct mammary gland expressing vectors. The DNA/liposome complex was injected into mammary glands of pregnant rabbit through duct of mammary gland. The results showed that both of the two vectors could efficiently express tPA with the activities of lysis of fibrin clot in the milk. The expressing quantity was between 200-500ng/ml. These results showed that these two vectors could be expressed in cells of mammary gland in high level and they could be used to establish transgenic animals. It also suggested that the direct injection of gene into mammary gland may become a simple and efficient method to produce proteins with biological activities.
出处 《高技术通讯》 EI CAS CSCD 1999年第10期52-56,共5页 Chinese High Technology Letters
基金 863计划资助
关键词 人组织型 纤溶酶原激活剂 载体 乳腺定位表达 Tissue plasminogen activator, Expression, Milk, Rabbit
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参考文献11

  • 1韩素文 俞炜源 等.-[J].军事医学科学院院刊,1987,11:101-118.
  • 2岳军明.中国人民解放军农牧大学博士论文[M].,1996.40.
  • 3张克中.-[J].生物工程学报,1997,13:419-419.
  • 4J.萨姆布鲁克.分子克隆实验指南,第二版[M].科学出版社,1989..
  • 5欧阳应斌.-[J].生物技术通讯,1993,4:163-163.
  • 6张克中,生物工程学报,1997年,13卷,419页
  • 7岳军明,博士学位论文,1996年
  • 8欧阳应斌,生物技术通讯,1993年,4期,163页
  • 9金冬雁(译),分子克隆实验指南(第2版),1989年
  • 10韩素文,军事医学科学院院刊,1987年,11卷,101页

共引文献1

同被引文献10

  • 1孙树权 张平武 戴建新 等.基因工程原理与方法[M].北京:人民军医出版社,2001.251.
  • 2Qiu X, Lu D, Zhou J, et al. Implantation of autologous skin fibroblast genetically modified to secrete clotting factor Ⅸ partially corrects the hemorrhagic tendencies in two hemophilia B patients[J]. Chin Med J(Engl), 1996,109(11):832-839.
  • 3Rosnoblet C, Vischer UM,Gerard RD,et al. Storage of tissue-type plasminogen activator in Weibel-Palade bodies of human endothelial cells[J]. Arterioscler Thromb Vasc Biol, 1999, 19(7):1796-1803.
  • 4Gibson CM, Cannon CP, Murphy SA, et al. Weight-adjusted dosing of TNK-tissue plasminogen activator and its relation to angiographic outcomes in the thrombolysis in myocardial infarction 10B trial. TIMI 10B Investigators[J].Am J Cardiol, 1999,84(9):976-980.
  • 5Waugh JM, Kattash M, Li J, et al. Gene therapy to promote thromboresistance: Local over expression of tissue plasminogen activator to prevent arterial thrombosis in an in vivo rabbit model[J]. Proc Natl Acad Sci USA, 1999,96(3):1065-1070.
  • 6Gibson CM, Cannon CP, Murphy SA, et al. Weight-adjusted dosing of TNK-tissue plasminogen activator and its relation to angiographic outcomes in the thrombolysis in myocardial infarction 10B trial. TIMI 10B Investigators[J].Am J Cardiol,1999,84(9):976-980
  • 7Waugh JM, Kattash M, Li J,et al. Gene therapy to promote thromboresistance: Local over expression of tissue plasminogen activator to prevent arterial thrombosis in an in vivo rabbit model[J]. Proc Natl Acad Sci USA, 1999,96(3):1065-1070
  • 8Rosnoblet C, Vischer UM,Gerand RD,et al. Storage of tissue-type plasminogen activator in Weibel-Palade bodies of human endothelial cells[J]. Arterioscler Thromb Vasc Biol 1999, 19(7):1796-1803
  • 9赵永波,李钰,张贵寅,关振中.携带tPA基因转移的真核表达载体的构建研究[J].中国地方病学杂志,1999,18(6):401-403. 被引量:2
  • 10陈勇,陈玉祥,龙志高,赵迪诚,潘乾,戴和平,夏昆,夏家辉.皮肤成纤维细胞体外培养的安全性实验研究[J].中国医师杂志,2002,4(11):1176-1178. 被引量:7

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