Abstract

Sweet tasting and taste modifying proteins are the natural alternative to flavor enhancers and artificial sweeteners. Curculin is a unique protein, which has both sweet tasting and taste modifying properties. In this study, pinII Ti plasmid vector was constructed with curculin gene, where the gene encoding curculin protein was placed under the control of the duel cauliflower mosaic virus 35S (2× CaMV 35S) promoter into rice (Oryza sativa L. var. Japonica cv. Dongjinbyeo) by Agrobacterium-mediated transformation to generate transgenic plants. Here, twenty-six plant lines were regenerated and the transgenic rice plant lines were confirmed by different molecular analysis. The genomic PCR result revealed that all of the plant lines were transgenic. The single copied and intergenic plant lines were selected by Taqman PCR analysis and FST analysis, respectively. Expression of curculin gene in transgenic rice resulted in the accumulation of curculin protein in the leave. The transgenic lines obtained in next generation, where curculin protein was also accumulated in the plant leave. Sensory analysis result suggested that the curculin protein expressing transgenic lines exerted both sweet and taste modifying activities. These results demonstrated that curculin was expressed in transgenic rice plants.

中文翻译：

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Curculin，一种新型替代甜味剂在转基因水稻中的表达

摘要

甜味和调味蛋白是增味剂和人造甜味剂的天然替代品。仙茅素是一种独特的蛋白质，具有甜味和增味特性。在这项研究中，用curculin基因构建了pinII Ti质粒载体，其中将编码curculin蛋白的基因置于对花椰菜花叶病毒35S（2×CaMV 35S）启动子的控制下进入水稻（Oryza sativa L. var。Japonica cv）。 。Dongjinbyeo）通过土壤杆菌介导的转化以产生转基因植物。在此，再生了二十六个植物品系，并且通过不同的分子分析证实了转基因水稻植物品系。基因组PCR结果显示所有植物系都是转基因的。通过Taqman PCR分析和FST分析分别选择单拷贝和基因间的植物品系。仙茅素基因在转基因水稻中的表达导致仙茅中仙茅素蛋白的积累。下一代获得的转基因品系，其中仙茅素蛋白也积累在植物叶片中。感官分析结果表明，表达仙茅素蛋白的转基因株系具有甜味和味觉调节活性。这些结果证明仙茅素在转基因水稻植物中表达。