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  

PCR-mediated modification of DFR gene from ornamental sunflower and construction of the gene′s plant expression vector

作　　者：张剑亮[1] 潘大仁[1] 周以飞[1] 吴明伟[1] 柯祥德[1] 林钿[1] 

机构地区：[1]福建农林大学作物科学学院,福建福州350002

出　　处：《福建农林大学学报（自然科学版）》2008年第2期166-169,共4页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：福建省自然科学基金资助项目(2007J0055);福建省教育厅资助项目(JB04307)

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.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.

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 

分 类 号：S565.5[农业科学—作物学] Q781[生物学—分子生物学]