Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of Ukraine, Zabolotnogo str 148, Kyiv 03680, Ukraine.
Plant Cell Rep. 2011 Mar;30(3):407-15. doi: 10.1007/s00299-010-0942-5. Epub 2010 Nov 3.
In this study, we report the obtaining of carrot plants expressing human interferon alpha-2b via Agrobacterium-mediated transformation using two vector constructs containing the sequence coding for interferon gene fused with Nicotiana plumbagenifolia calreticulin apoplast targeting signal driven by 35S CaMV promoter and root-specific Mll promoter. The human interferon alpha-2b gene was correctly translated in carrot plants according to Western blot analysis. The recombinant protein exhibited antiviral activity in vitro by inhibition of vesicular stomatitis virus replication in established piglet testicular cells. The results demonstrated the higher activity of interferon accumulated in carrot plants for young leaves (up to 50.7 × 10(3) IU/g FW) compared to the mature ones probably due to the degradation-susceptible nature of this protein. The taproot-expressing system could have also provided the sufficient protein amounts (up to 16.5 × 10(3) IU/g FW) and could possibly be used for generating interferon alpha-2b protein in planta for preventing and curing infectious diseases.
在这项研究中，我们通过利用含有与人干扰素α-2b 基因序列融合的两个载体构建体，报告了通过农杆菌介导的转化获得表达人干扰素α-2b 的胡萝卜植株，该序列融合了 Nicotiana plumbagenifolia calreticulin 质外体靶向信号，由 35S CaMV 启动子和根特异性 Mll 启动子驱动。根据 Western blot 分析，人干扰素α-2b 基因在胡萝卜植株中正确翻译。重组蛋白在体外通过抑制已建立的猪睾丸细胞中的水疱性口炎病毒复制表现出抗病毒活性。结果表明，与成熟叶片相比，干扰素在幼叶中的积累活性更高（高达 50.7×10^3 IU/g FW），这可能是由于该蛋白易降解。根表达系统也可能提供了足够的蛋白量（高达 16.5×10^3 IU/g FW），并可能用于在植物体内生成干扰素α-2b 蛋白，以预防和治疗传染病。
