果实特异性RNAi介导的Lcy基因沉默来增加番茄中番茄红素的含量
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Fruit-specific RNAi-mediated Lcy Gene Silencing Enhances Content of Lycopene in Tomatoes
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    摘要:

    根据GenBank中番茄的番茄红素β-环化酶(Lcy)基因序列和八氢番茄红素去饱和酶基因(Pds)启动子序列设计特异引物从番茄基因组DNA中分别扩增出了Lcy基因的高度保守的长302bp的DNA片段和长1790的Pds启动子片段。根据RNAi的原理,将Lcy基因的DNA片段以正反两个方向通过一段内含子序列连接在一起形成RNAi片段,将该片段与Pds启动子一起插入到pVCT2020的表达载体中,通过农杆菌介导的方法转化番茄,获得转基因植株5棵,PCR检测证实外源片段已成功导入番茄基因组中。收获转色期后20d左右的完全成熟的番茄果实提取番茄红素进行含量分析,结果显示:转基因番茄果实中番茄红素的含量极大的增加了。上述结果表明:通过RNAi果实特异性的抑制类胡萝卜素代谢途径中生物合成酶基因的表达能够极大的增加番茄果实中番茄红素的含量。这为通过基因工程手段提高番茄果实中的营养价值提供了参考

    Abstract:

    Tomatoes (Lycopersicon esculentumMill.) are the principal dietary source of Lycopene which is one of carotenoid and is highly beneficial in preventing some diseases such as the cancer and the heart disease. Suppressing the expression ofLcy gene,the main gene regulating the transformation of the lycopene ,is a convenient and effective way to enhance the content of lycopene. The primers were designed according to the gene sequence(U46919)and (X86452) in GenBank. The fruit-specific promoter—phytoene desaturase gene(Pds) promoter and the DNA segment of the Lcy gene were isolated from the genome DNA of tomatoes. The 3′end ofLcy DNA segment was connected together by an intron to inform the RNA interferential segment then which was inserted in the expression vector with the Pdspromoter to inform the fruit-specific expression vector. The vector was transformed into the tomatoes through theAgrobacterium tumefaciens. Five transformants were obtained. And the PCR proved that the extra_gene was integrated into the tomato genome. The lycopene in the transgenic tomatoes fruit was increased significantly through analysing the contents of lycopene. These results shows that regutating biosynthetic enzyme in carotenoid pathway by RNAi can improve the lycopene content of plant-derived products.

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万群,张兴国,宋明. 果实特异性RNAi介导的Lcy基因沉默来增加番茄中番茄红素的含量[J]. 生物工程学报, 2007, 23(3):

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