| 加强Psy基因表达和通过RNAi抑制Lcy基因表达的载体构建 |
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| 引用本文： | 万群,张兴国,宋明,苏承刚.加强Psy基因表达和通过RNAi抑制Lcy基因表达的载体构建[J].农业生物技术学报,2007,15(3):477-481. |
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| 作者姓名： | 万群 张兴国 宋明 苏承刚 |
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| 作者单位： | 西南大学园艺园林学院,重庆市蔬菜学重点实验室,重庆,400716 |
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| 摘 要： | 通过基因工程手段加强八氢番茄红素合成酶（Psy）基因的表达和通过RNAi抑制番茄红素β-环化酶（Lcy）基因（该基因是调控番茄红素转化为其他类胡萝卜素的主要的基因）的表达是培育高番茄红素品种的重要手段之一。根据GenBank中拟南芥的Psy基因序列（NM-121729），通过聚合酶链式反应（PCR）从拟南芥的cDNA中扩增出了长1296的Psy基因。根据GenBank中番茄的Lcy基因序列（X86452）和Pds启动子序列（U46919）分别设计特异引物从番茄中扩增出了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基因的长302bp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片段插入到抗Kan的pVCT2020的表达载体中，并通过具有果实特异性特点的启动子——八氢番茄红素去饱和酶基因（Pds）启动子控制该干扰片段的表达，同时将Psy基因在Pds启动子调控下插入到同一载体中，从而构建成了能同时加强Psy基因的表达和抑制Lcy基因表达的植物表达载体。
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| 关 键 词： | 番茄 Psy基因 Lcy基因 Pds启动子 |
| 文章编号： | 1006-1304（2007）03-0477-05 |
| 收稿时间： | 2006-09-21 |
| 修稿时间： | 2006-09-212006-10-24 |
Construction of Expression Vector for Enhancing Psy Gene Expression and Blocking Lcy Gene Expression through RNAi |
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| WAN Qun,ZHANG Xing-guo,SONG Ming,SU Cheng-gang.Construction of Expression Vector for Enhancing Psy Gene Expression and Blocking Lcy Gene Expression through RNAi[J].Journal of Agricultural Biotechnology,2007,15(3):477-481. |
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| Authors: | WAN Qun ZHANG Xing-guo SONG Ming SU Cheng-gang |
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| Institution: | Key Laboratory in Olericulture of Chongqing, College of Horticulture and Landscape, Southwest University,Chongqing 400716, China |
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| Abstract: | Enhancing the expression of phytoene synthase (Psy )gene and blocking the expression of lycopene cyclase (Lcy )gene, which are the main genes regulating the transformation of the lycopene,are the convenient way to increase the content of lycopene. The primers were designed according to the gene sequence NM-121729, U46919 and X86452 in GenBank. The Psy gene was isolated from the cDNA of the Arabidopsis thaliana. The phytoene desaturase gene(Pds ) promoter and the DNA segments of the Lcy gene were obtained from the genome DNA of tomato(Lycopersicon esculentum Mill.). The 3'end of Lcy DNA segment was connected together with an intron to inform the RNA interference (RNAi) segment and inserted into the expression vector with the fruit-specific promoter(Dds );the Psy gene was also inserted into the same vector under the manipulation of the Pds promoter,then the expression vector was obtained and could enhance the expression of Psy gene and interfere the expression of Lcy gene at the same time by transforming it into the plant. |
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| Keywords: | RNAi |
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