玉米淀粉分支酶基因片段的克隆和植物表达载体的构建

Cloning and Construction of Expression Vector on Gene of Corn Starch Branching Enzyme

应用聚合酶链式反应技术(PCR)扩增了玉米淀粉分支酶的cDNA基因片段,并将其克隆到pMD18-Tvector载体上,对重组子进行了PCR检测和限制性内切酶分析,测定了DNA全序列。结果表明:克隆片段全长为935bp。将反义+正义基因片段插入到pBI12135S启动子下,构建成表达质粒pCJSBE2b。通过花粉管通道法将其导入玉米自交系,获得了转基因植株。

<Abstrcat> cDNA segment of corn starch branching enzymes gene was obtained by PCR technique and cloned into pMD18-Tvector. The recombinant clone was detected by PCR technique and analyzed by the restriction enzyme. A full length of cDNA gene was sequenced. The results showed that the length of the clone sequence was 935 bp. The fragment of anti-sense gene + plus-sense gene were inserted into 35S promoters of pBI121 respectively to construct expression plasmids pCJSBE2b. Then we transformed them into different corn inbred lines by using Pollen-tube Pathway. Finally we got transgenic plants.