Cheon Ban Yoon, Kim Hae Jin, Oh Kyung Hee, Bahn Sung Chul, Ahn Ji Hoon, Choi Jang Won, Ok Sung Han, Bae Jung Myung, Shin Jeong Sheop
School of Life Sciences and Biotechnology, Korea University, Seoul 136-701, Korea.
Transgenic Res. 2004 Dec;13(6):541-9. doi: 10.1007/s11248-004-2737-3.
Erythropoietin (EPO) is a glycoprotein used for curing human anemia by regulating the differentiation of erythroid progenitors and the production of red blood cells. To examine the expression of recombinant EPO in plants, pPEV-EP21, in which human epo cDNA under the control of the CaMV 35S promoter, was introduced into tobacco and Arabidopsis via Agrobacterium tumefaciens-mediated transformation. The RNA expression level of epo in the transgenic lines was initially estimated by Northern blot analysis. Two transgenic lines, which exhibited a high expression level of epo mRNA determined by Northern analysis, were chosen for Western blot analysis to examine the production of EPO proteins. Those two lines, EP21-12 and EP21-14, revealed detectable bands on the immunoblot. Interestingly, constitutive expression of the human epo gene affected the morphologies in transgenic plants such that vegetative growth of transgenic tobacco was retarded, and male sterility was induced in transgenic tobacco and Arabidopsis.
促红细胞生成素(EPO)是一种糖蛋白,通过调节红系祖细胞的分化和红细胞的生成来治疗人类贫血。为了检测重组EPO在植物中的表达,将在花椰菜花叶病毒35S启动子控制下的人促红细胞生成素cDNA的pPEV-EP21,通过根癌农杆菌介导的转化导入烟草和拟南芥中。通过Northern印迹分析初步估计转基因株系中促红细胞生成素的RNA表达水平。选择通过Northern分析确定促红细胞生成素mRNA表达水平高的两个转基因株系进行Western印迹分析,以检测EPO蛋白的产生。这两个株系,EP21-12和EP21-14,在免疫印迹上显示出可检测到的条带。有趣的是,人促红细胞生成素基因的组成型表达影响了转基因植物的形态,使得转基因烟草的营养生长受到抑制,并在转基因烟草和拟南芥中诱导了雄性不育。