| 甘蓝型油菜依赖焦磷酸的磷酸果糖激酶(PFK)基因片段克隆及hpRNAi载体构建 |
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| 引用本文： | 汪承刚,张菊红,谢妤,刘幼琪,黄邦全.甘蓝型油菜依赖焦磷酸的磷酸果糖激酶(PFK)基因片段克隆及hpRNAi载体构建[J].分子植物育种,2006,4(2):199-204. |
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| 作者姓名： | 汪承刚 张菊红 谢妤 刘幼琪 黄邦全 |
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| 作者单位： | 湖北大学生命科学学院植物遗传育种实验室,武汉,430062 |
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| 基金项目： | 湖北省科技厅国际合作项目;湖北省教育厅国际合作项目 |
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| 摘 要： | 依赖焦磷酸的磷酸果糖激酶(PFK)是糖酵解途径的关键酶。本研究首先构建带有种子特异性napin启动子和Nos终止子的植物表达载体2300-nap及带有组成型35S启动子和Nos终止子的植物表达载体2300-35S;然后用PCR法从甘蓝型油菜(BrassicanapusL.)中油119总基因组中扩增出依赖焦磷酸的磷酸果糖激酶(PFK)基因片段,再以扩增出的PFK基因片段作模板设计引物扩增出一个相应的小片段。将两个PFK基因片段反向连接,插入到植物表达载体2300-nap的napin启动子和nos终止子之间,植物表达载体2300-35S的35S启动子和nos终止子之间,分别构建成可转录表达出发夹RNA(hairpinRNA,hpRNA)结构的种子特异型和组成型油菜RNA干扰载体,为今后油菜利用RNA干扰(RNAi)提高含油量的基因工程研究奠定了基础。
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| 关 键 词： | 甘蓝型油菜 依赖焦磷酸的磷酸果糖激酶(PFK) 发夹RNA(hpRNA) 载体构建 |
Cloning and hpRNAi Vector Construction of Brassica napus Pyrophosphate-dependent Phosphofructokinase (PFK) |
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| Wang Chenggang,Zhang Juhong,Xie Yu,Liu Youqi,Huang Bangquan.Cloning and hpRNAi Vector Construction of Brassica napus Pyrophosphate-dependent Phosphofructokinase (PFK)[J].Molecular Plant Breeding,2006,4(2):199-204. |
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| Authors: | Wang Chenggang Zhang Juhong Xie Yu Liu Youqi Huang Bangquan |
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| Abstract: | Pyrophosphate-dependent phosphofructokinase (PFK) plays an important role in the EMP pathway in rapeseed (Brassica napus L.). Firstly we constructed plant expression vector 2300-nap harboring seed-specific napin promoter and nos terminator, and plant expression vector 2300-35S harboring constitutive 35S promoter and nos terminator. PFK gene fragment was PCR-amplified from B. napus genomic DNA and was used as template to amplify a smaller fragment. These two PFK fragments were ligated in opposite directions into plant expression vector 2300-nap and 2300-35S respectively to produce hpRNAi vector hp-PFK-nap and hp-PFK-35S. This is the first step to increase oil content in rapeseed using RNA interference. |
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| Keywords: | Brassica napus Pyrophosphate-dependent phosphofructokinse (PFK) hpRNA Vector construction |
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