| 番茄叶片GDP-甘露糖焦磷酸化酶基因表达载体的构建与转化 |
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| 引用本文： | 吴海琴,赵瑛,符庆功,王华森.番茄叶片GDP-甘露糖焦磷酸化酶基因表达载体的构建与转化[J].生物技术通报,2010(2). |
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| 作者姓名： | 吴海琴 赵瑛 符庆功 王华森 |
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| 作者单位： | 浙江林学院农业与食品科学学院,临安,311300 |
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| 摘 要： | GDP-甘露糖焦磷酸化酶(GMPase,EC 2.7.7.22)是维生素C合成途径的第一步关键酶。通过RT-PCR扩增到1498 bp的GMPase全长序列,GenBank登录号为DQ449030。利用克隆到的基因分别构建得到正义及反义植物表达载体。并将其克隆至PBI121真核表达载体,转化农杆菌LBA4404,利用农杆菌介导法转化烟草植株,经基因组PCR及琼脂糖凝胶电泳检测,结果表明GMPase真核表达载体构建成功,并成功获得转基因植株。
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| 关 键 词： | 番茄 GDP-甘露糖焦磷酸化酶基因(GMPase) 表达载体 |
Construction of GDP-mannose Pyrophosphorylase Gene Expression Vector and Western Blotting |
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| Wu Haiqin,Zhao Ying,Fu Qinggong,Wang Huasen.Construction of GDP-mannose Pyrophosphorylase Gene Expression Vector and Western Blotting[J].Biotechnology Bulletin,2010(2). |
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| Authors: | Wu Haiqin Zhao Ying Fu Qinggong Wang Huasen |
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| Institution: | Wu Haiqin Zhao Ying Fu Qinggong Wang Huasen(Hangzhou Department of Horticulture,School of Agricultura , Food Science,Zhejiang Forestry University,Linan 311300) |
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| Abstract: | GDP-mannose pyrophosphorylase(GMPase,EC 2.7.7.22)can catalyse the synthesis of GDP-D-mannose and represents the first committed step in the formation of all guanosin-containing sugar nucleotides found in plants which are precursors for the synthesis of L-ascorbate.A full-length cDNA encoding GMPase from Lycopersicon esculentum was isolated using a RT-PCR.Transgenic tobacco plants were generated in which the fragments of tomato GDP-mannose pyrophosphorylase gene(GMPase)cDNA was introduced in antisense orient... |
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| Keywords: | AsA Tomato GDP-mannose pyrophosphorylase AsA Expression vector |
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