自身启动子驱动的白菜型油菜FAE1家族RNAi载体构建

Construction of RNAi Vector of Brassia campestris FAE1 Gene Family Drive by Autologous Promoter

白菜型油菜具有许多其他油菜所不具有的优点,如耐旱、抗寒能力强、生长周期短等,但较为突出的缺点是芥酸在其脂肪酸中的含量较高,并且通过常规育种方法很难获得低芥酸的品种。pFGC5941M载体是由pFGC5941载体改进而来,它含有多个酶切位点,是较为理想的载体骨架,但它的CaMV 35S启动子是组成型的强表达启动子,不具备特异性,不利于我们有目的地观察某基因的功能。本研究克隆了白菜型油菜FAE1家族两个成员的启动子片段和FAE1 RNA干扰片段,并用FAE1的启动子代替了植物表达载体pFGC5941M的组成型表达启动子CaMV 35S,构建了自身启动子驱动的FAE1 RNAi载体,为通过基因工程手段降低白菜型油菜种子中芥酸的含量,提高其油用价值提供基础。

Compared with other rape, Brassia campestris had many advantages, such as high drought, cold resistance and short growth cycle. But it has a prominent disadvantage, which had high erucic acid content, and it is hard to cultivate low erucic acid breed by conventional breeding, pFGC5941M was modified by pFGC5941, which contained multiple restriction enzyme sites and was an ideal vector. But its CaMV 35S promoter was a strong expression promoter and didn＇t conducive to study the function of a particular gene. In this study, two promoters and RNA interference fragment of FA E1 gene family were cloned. We used FA E1 promoters instead of CaMV 35S promoter in pFGC5941M and constructed RNAi vector ofBrassia campestris FAE1 gene family drive by autologous promoter. This is the basis to reduce the erucic acid content in seeds by genetic engineering and improve oil value ofBrassia campestris.