| 甘蓝型油菜乙醇酸氧化酶(GO)基因片段的克隆及hpRNA表达载体构建 |
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| 引用本文： | 汪承刚,李长春,谢妤,刘幼琪,黄邦全.甘蓝型油菜乙醇酸氧化酶(GO)基因片段的克隆及hpRNA表达载体构建[J].湖北大学学报(自然科学版),2007,29(3):286-289,324. |
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| 作者姓名： | 汪承刚 李长春 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 湖北武汉430062 |
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| 基金项目： | 湖北省科技厅国际合作重点项目(2005CA020)，湖北省教育厅国际合作项目(G200510001)资助 |
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| 摘 要： | 通过PCR从甘蓝型油菜(Brassica napus)华双4号基因组DNA中扩增出乙醇酸氧化酶(GO)基因片段,DNA序列分析表明扩增GO基因片段的外显子部分与报道序列相同.以扩增出的GO基因片段作模板从一端设计引物扩增出一个相应的小片段.将GO基因大小两个片段反向连接,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(hairpin RNA,hpRNA)结构的种子特异型和组成型油菜hpRNA干扰载体.
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| 关 键 词： | 甘蓝型油菜 乙醇酸氧化酶(GO) 光呼吸 净光合速率 发夹RNA(hpRNA) 载体构建 |
| 文章编号： | 1000-2375（2007）03-0286-04 |
| 收稿时间： | 2006-09-18 |
| 修稿时间： | 2006-09-18 |
Cloning and hpRNA vector construction for Brassica napus glycolate oxidase gene (GO) |
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| WANG Cheng-gang,LI Chang-chun,XIE Yu,LIU You-qi,HUANG Bang-quan.Cloning and hpRNA vector construction for Brassica napus glycolate oxidase gene (GO)[J].Journal of Hubei University(Natural Science Edition),2007,29(3):286-289,324. |
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| Authors: | WANG Cheng-gang LI Chang-chun XIE Yu LIU You-qi HUANG Bang-quan |
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| Institution: | School of Life Science, Hubei University, Wuhan 430062,China |
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| Abstract: | Glycolate oxidase(GO) is a key enzyme for plant photorespiration.GO Gene fragment was PCR-amplified from B.napus genomic DNA and then used as template to amplify a smaller fragment.The larger and the smaller fragments were ligated in opposite orientations into a seed specific expression vector 2300-nap and constitutive vector 2300-35S respectively to produce hpRNAi vector hp-GO-nap and hp-GO-35S.Sequence analysis indicated that the GO fragment we amplified had 100% similarity in the exon part with the published sequence in the Genbank. |
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| Keywords: | Brassica napus glycolate oxidase(GO) photorespiration hpRNA vector construction |
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