蒺藜苜蓿二氢黄酮还原酶基因克隆及植物表达载体构建

Cloning and construction of plant expression vector of dihydroflavonol reductase gene from Medicargo truncatula

采用RT-PCR方法,从蒺藜苜蓿(Medicargotrunctula)中克隆了二氢黄酮还原酶(DFR)基因cDNA片段,并进行DFR基因植物表达载体的构建.结果表明:扩增的DFR基因片段全长约1 000 bp,编码337个氨基酸,与Gen-Bank中已注册的DFR基因核苷酸序列的同源性为99.80%.以pBI121为基础载体,构建了CaMV35S启动子驱动的DFR基因的植物表达载体pBIDFR,然后导入农杆菌EHA105,经PCR验证确定构建出植物表达载体.

In this study,RT-PCR was employed to clone cDNA fragment of dihydroflavonol reductase（DFR） gene of 1 000 bp from Medicargo truncatula;Sequence analysis showed that 99.8 % of the DFR were homologous to those in GenBank.And this sequence encoded 337 amino-acid residues.Plant expression vector named pBIDFR was constructed based on pBI121 vector.Moreover,pBIDFR was transferred into Agrobacterium tumefacien EHA105 by direct DNA transfer.