Expression of two subtypes of human IFN-α in transgenic potato plants
Expression of two subtypes of human IFN-α in transgenic potato plants
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DOI：
10.1089/10799900152547858
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发表时间：
2001-08-01
影响因子：
2.3
通讯作者：
Sugimoto, C
中科院分区：
文献类型：
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作者：
Ohya, K;Matsumura, T;Sugimoto, C
植物表达系统在低生产成本和低动物病毒或细菌内毒素污染风险方面具有优于其他体外表达系统的优势。在这项研究中，cDNA编码的两个亚型的人干扰素-α 2b和8（HuIFN-α 2b和HuIFN-α 8）被引入到马铃薯植物（Solanum tuberosum）使用农杆菌介导的转化。分别通过北方印迹分析和ELISA证实插入的HuIFN-α cDNA的转录和翻译。通过抑制水泡性口炎病毒（VSV）在人羊膜细胞系上的复制来测定产物的生物活性。然而，由于马铃薯组织提取物中存在对动物细胞有毒的物质，只有在通过透析除去这些物质后才能成功证明转化体中的IFN生物活性。植物提取物中IFN活性的最高水平为560 IU/g组织。这些结果表明，导入马铃薯植物的HuIFN-α基因在植物细胞中正确翻译和转录。这一报道首次表明具有潜在医药应用价值的具有生物活性的动物细胞因子可以在转基因马铃薯植物中表达。
Plant expression systems have advantages over other in vitro expression systems in terms of low production costs and low risk of contamination by animal viruses or bacterial endotoxins. In this study, cDNA encoding two subtypes of human interferon-alpha 2b and 8 (HuIFN-alpha 2b and HuIFN-alpha8) were introduced into potato plants (Solanum tuberosum) using Agrobacterium-mediated transformation. Transcription and translation of the inserted HuIFN-alpha cDNA were confirmed by Northern blot analysis and ELISA, respectively. Bioactivity of the products was assayed by inhibition of vesicular stomatitis virus (VSV) replication on a human amniotic cell line. However, because of the presence of substances in potato tissue extracts that were toxic to animal cells, successful demonstration of IFN bioactivity in the transformants; was achieved only after removal of such substances by dialysis. The maximum level of IFN activity in plant extracts was 560 IU/g of tissue. These results indicated that the HuIFN-alpha gene introduced into the potato plant was correctly translated and transcribed in plant cells. This report for the first time shows that biologically active animal cytokines with potential pharmaceutical applications could be expressed in transgenic potato plants.