| PCR介导观赏向日葵DFR基因改造及其真核表达载体构建研究初报 |
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| 引用本文： | 张剑亮,潘大仁,周以飞,吴明伟,柯祥德,林钿.PCR介导观赏向日葵DFR基因改造及其真核表达载体构建研究初报[J].福建农林大学学报(自然科学版),2008,37(2):166-169. |
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| 作者姓名： | 张剑亮 潘大仁 周以飞 吴明伟 柯祥德 林钿 |
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| 作者单位： | 福建农林大学作物科学学院,福建,福州,350002 |
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| 基金项目： | 福建省自然科学基金
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福建省教育厅科研项目 |
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| 摘 要： | 二氢黄酮醇-4-还原酶(DFR)是花色素苷代谢途径中的关键酶之一.本研究利用PCR技术将观赏向日葵DFR底物结合区敲除,并将该区域中134位保守氨基酸天冬酰胺定点突变为天门冬氨酸和亮氨酸,最后获得已敲除底物结合区的基因KODFR以及定点突变的基因N134D和N134L,并成功构建KODFR、N134D和N134L基因的植物表达载体pCAM-KODFR、pCAM-N134D和pCAM-N134L.
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| 关 键 词： | 观赏向日葵 PCR 二氢黄酮醇-4-还原酶 底物结合位点突变 载体构建 |
| 文章编号： | 1671-5470(2008)02-0166-04 |
| 修稿时间： | 2007年11月10 |
PCR-mediated modification of DFR gene from ornamental sunflower and construction of the gene's plant expression vector |
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| ZHANG Jian-liang,PAN Da-ren,ZHOU Yi-fei,WU Ming-wei,KE Xiang-de,LIN Dian.PCR-mediated modification of DFR gene from ornamental sunflower and construction of the gene's plant expression vector[J].Journal of Fujian Agricultural and Forestry University,2008,37(2):166-169. |
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| Authors: | ZHANG Jian-liang PAN Da-ren ZHOU Yi-fei WU Ming-wei KE Xiang-de LIN Dian |
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| Abstract: | Dihydroflavonol 4-reductase(DFR) catalyzes the last common step in the anthocyanin biosynthesis pathway.By using PCR,substrate binding region of DFR from ornamental sunflower(Helianthus annuus) was knocked out,and-134 conserved amino acid residue asparagine was mutated into aspartate or leucine.KODFR,mutated N134D and N134L were successfully amplified.Plant expression vector pCAM-KODFR,pCAM-N134D and pCAM-N134L were also successfully constructed,which will be used for gene transformation. |
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| Keywords: | ornamental sunflower PCR dihydroflavonol 4-reductase mutation of substrate binding region vector construction |
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