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利用TMV病毒载体在烟草中瞬时表达rPA(Reteplase)
引用本文:雒丽娜,王盛.利用TMV病毒载体在烟草中瞬时表达rPA(Reteplase)[J].宁夏大学学报(自然科学版),2012,33(1):58-61.
作者姓名:雒丽娜  王盛
作者单位:1. 宁夏大学生命科学学院,宁夏银川,750021
2. 宁夏大学生命科学学院,宁夏银川 750021;宁夏大学西部特色生物资源保护与利用教育部重点实验室,宁夏银川 750021
基金项目:国家自然科学基金资助项目(31160032)
摘    要:构建了携带有人组织型纤溶酶原激活剂突变体rPA(Reteplase)的烟草花叶病毒(Tobacco Mosaic Virus,TMV)瞬时表达载体PJL TRBO-rPA,并转入农杆菌GV3101中.利用农杆菌渗滤接种技术在烟草本氏烟(Nicotiana benthamiana)中进行表达.经SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)、免疫印迹(western blotting)和纤维蛋白琼脂糖平板(fibrin-agarose plate assay,FAPA)法检测分析.结果表明,在烟草中成功地表达出了有活性的rPA.为在植物中生产Reteplase提供了一条可能的途径.

关 键 词:人组织型纤溶酶原激活剂突变体rPA(Reteplase)  烟草花叶病毒  瞬时表达载体

Expression of the Truncated Human Tissue Plasminogen Activator (Reteplase) in Plant Leaves Using Tobacco Mosaic Virus RNA-Based Expression System
Luo Lina , Wang Sheng.Expression of the Truncated Human Tissue Plasminogen Activator (Reteplase) in Plant Leaves Using Tobacco Mosaic Virus RNA-Based Expression System[J].Journal of Ningxia University(Natural Science Edition),2012,33(1):58-61.
Authors:Luo Lina  Wang Sheng
Institution:1,2*(1.School of Life Science,Ning xia University,Yinchuan 750021,China; 2.Key Laboratory of Ministry of Education for Protection and Utilization of Special Biological Resources in the Western China,Ningxia University,Yinchuan 750021,China)
Abstract:Tobacco Mosaic Virus RNA-based transient expression vectors containing the truncated Human Tissue Plasminogen Activator(Reteplase) was constructed and transformed into Agrobacterium GV3101.The recombinant Agrobacteria were delivered into tobacco plants(Nicotiana benthamiana) by means of an Agrobacterium-mediated transient transformation assay.The efficiency was evaluated by SDS-PAGE,western blotting and Fibrin-agarose plate assay(FAPA).Those results showed that bio-active rPA was successfully expressed in the tobacco leaves and proposed that the plant virus RNA-based expression system will be a useful tool for expressing Reteplase in plants for production purposes.
Keywords:Human Tissue Plasminogen Activator(Reteplase)  Tobacco Mosaic Virus  RNA-based transient expression vector
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