OBTAINING OF THE TRANSGENIC HELIANTHUS TUBEROSUS L. PLANTS, CALLUS AND "HAIRY" ROOT CULTURES ABLE TO EXPRESS THE RECOMBINANT HUMAN INTERFERON ALPHA-2b GENE.

TSitologiia i genetika Pub Date : 2015-09-01
O M Maistrenko, Yu S Luchakivska, N M Zholobak, M Ya Spivak, M V Kuchuk
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Abstract

This work is the first to our knowledge to describe the successful attempt of Agrobacterium rhizogenes-mediated transformation of topinambour in order to obtain the transgenic H. tuberosus plants, callus and "hairy" root cultures. The plasmid vectors contained the sequence of interferon gene fused with Nicotiana plumbagenifolia L. calreticulin apoplast targeting signal driven by 35S CaMV promoter or root-specific Mll promoter. Nearly 75% isolated Ri-root lines and callus cultures were proved (by PCR analysis) to contain HuINFa-2b transgene. We also managed to obtain H. tuberosus transgenic plants through somatic embryogenesis on the transgenic "hairy" root culture. The obtained transgenic H. tuberosus cultures exhibited high-level antiviral activity that ranged from 2000 to 54500 IU/g FW that makes this crop considered a promising source of recombinant interferon alpha 2b protein.

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获得能够表达重组人类干扰素 ALPHA-2b 基因的转基因 HELIANTHUS TUBEROSUS L. 植物、胼胝体和 "毛状 "根培养物。
据我们所知,这项研究首次成功地描述了根瘤农杆菌介导的topinambour转化,从而获得转基因H. tuberosus植株、胼胝体和 "多毛 "根培养物。质粒载体包含由 35S CaMV 启动子或根部特异性 Mll 启动子驱动的干扰素基因序列与 Nicotiana plumbagenifolia L. calreticulin apoplast targeting signal 融合。近 75% 分离出的 Ri-root 株系和胼胝体培养物被证明(通过 PCR 分析)含有 HuINFa-2b 转基因。我们还设法通过转基因 "多毛 "根培养物上的体细胞胚胎发生获得了转基因 H. tuberosus 植物。所获得的转基因 H. tuberosus 培养物表现出高水平的抗病毒活性,从 2000 IU/g FW 到 54500 IU/g FW 不等,这使得该作物被认为是重组干扰素 alpha 2b 蛋白的一个有前途的来源。
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