白菜型油菜BrDAD1基因的克隆及其植物反义载体的构建

Cloning and Construction of Plant Antisense Expression Vector Containing BrDAD1 Gene in Brassica rapa

应用聚合酶链式反应技术(PCR)扩增了白菜型油菜BrDAD1基因及自身的启动子基因Pro-DAD1,并将其分别克隆到pMD18-Tvector载体上,对重组子进行PCR检测和限制性内切酶分析,并测定了该基因及启动子全序列。结果表明,该基因及启动子全长分别为1 270 bp与1 335 bp。将白菜型油菜BrDAD1基因反向克隆到植物表达载体pCambia 1300的自身启动子基因Pro-DAD1启动子下游,构建了该基因的植物反义表达载体pCamDAD1。

BrDAD1 gene and its promoter Pro -DAD1 in Brassica rapa were obtained by PCR technique and cloned into pMD18 -Tvector respectively. The recombinant clone were detected by PCR technique and analyzed by the restriction enzyme. Two full lengths of BrDAD1 and Pro - DAN1 were sequenced. The results showed that the lengths of the clone sequences were 1 270 and 1335 bp respectively. BrDAD1 gene in Brassica rapa was reversely cloned into its promoter Pro-DAN1 downstream of plant expression vector pCambia 1300 in orientation, and then anti- sense expression vector pCamDAD1 has been constructed.