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

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.