棉花PEPC基因种子特异性ihpRNA表达载体的构建及鉴定

Construction and Identification of a Seed-specific Expression ihpRNA Vector Targeting the PEPC Gene of Cotton

磷酸烯醇式丙酮酸羧化酶(phosphoenolpyruvate carboxylase,PEPC)是控制植物体中蛋白质和脂肪酸含量比例的关键酶。本研究从棉花中克隆得到PEPC基因,长度为433bp,并将该基因的正反义片段分别和种子特异性启动子napin启动子(1123bp)、α球蛋白B基因启动子(1149bp)连接,插入到植物表达载体pCADS1341中。经酶切和PCR鉴定,成功的构建了PEPC基因的种子特异性ihpRNA表达载体pCADSNPSPA和pCADSBPSPA,为后期高含油量棉花材料的选育打下了基础。

Phosphoenolpyruvate carboxylase （PEPC） is a key enzyme, which controls the ratio of the protein/lipid content in plant. In this research, we cloned PEPC gene from cotton, which was 433 bp fragment, and the sense and antisense of PEPC gene were linked with seed-specific promoter of napin promoter and α globulin B gene promoter, which was 1 123 bp and 1 149 bp fragment respectively, then they were inserted into the plant expression vector of pCADS 1341. The enzyme digested and PCR amplified showed that the seed-specific ihpRNA vectors pCADSNPSPA and pCADSBPSPA were constructed successfully. This experiment would layed reference for breeding cotton materials with high oil content in the future.