Identification and characterization of highly expressed genes in suspension-cultured cells of sweet potato
Identification and characterization of highly expressed genes in suspension-cultured cells of sweet potato
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DOI：
10.1007/bf03178813
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发表时间：
2006-10-31
影响因子：
2.9
通讯作者：
Huh, Gyung Hye
中科院分区：
文献类型：
--
作者：
Kim, Young Hwa;Hur, Cheol Goo;Huh, Gyung Hye
培养的植物细胞在为大规模生产具有商业价值的重组蛋白提供高效且低成本的分子农业系统方面具有很大潜力。作为建立一种用于粮食作物悬浮培养细胞的高效表达系统的首要目标，我们首先从甘薯指数生长期细胞的cDNA文库中获得了1411个表达序列标签，并将它们组装成156个重叠群和1039个单拷贝序列。选择了5个表达序列标签作为最显著丰富的基因。它们分别是细胞壁发育蛋白（肉桂醇脱氢酶）、碳水化合物代谢蛋白（甘油醛 - 3 - 磷酸脱氢酶）和细胞周期调节蛋白（小GTP结合蛋白Ran）以及无机焦磷酸酶和亲环蛋白的转录本。然后与甘薯的根和叶的表达序列标签文库进行了比较。Northern杂交和逆转录 - 聚合酶链反应分析表明，这5个基因在悬浮细胞中强烈表达，但在根和叶中不表达，从而支持了从比较分析中获得的数据。这是首次对从同一植物物种的不同细胞类型中分离出的各种表达序列标签文库进行比较的报道。这些基因现在可有助于为从悬浮培养细胞设计一种高效表达系统提供适用的启动子。
Cultured plant cells have high potential in providing efficient and low-cost molecular farming systems for the large-scale production of commercially valuable recombinant proteins. As an initial aim at establishing an efficient expression system for suspension-cultured cells of food crops, we first obtained 1411 expressed sequence tags from a sweet potato cDNA library of exponential phase cells, and assembled them into 156 contigs and 1039 singletons. Five ESTs were selected as the most significantly abundant genes. These were transcripts for a cell wall development protein (cinnamyl alcohol dehydrogenase), a carbohydrate metabolite protein (glyceraidehyde-3-phosphate dehydrogenase), and a cell cycle regulator protein (small GTP binding protein Ran), as well as inorganic pyrophosphatase and cyclophilin. Comparisons were then made with the root and leaf EST libraries of sweet potato. Northern blot and RT-PCR analyses revealed that these five genes were strongly expressed in the suspension cells but not in the roots and leaves, thereby supporting the data obtained from the comparative analysis. This is the first reported comparison of the various EST libraries isolated from different cell types of the same plant species. These genes can now contribute to an applicable promoter for devising an efficient expression system from suspension-cultured cells.