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将水稻中等重复序列RRD3及其系列缺失体克隆到植物启动子检测载体中,通过根癌土壤杆菌介导转化水稻愈伤组织,利用GUS组织化学方法检测其在水稻愈伤组织中的启动子活性。结果显示:全长RRD3、410bp及150b缺失体具有强的启动子活性,而700bp、120bp缺失体仅有弱的启动子活性。通过与RRD3系列缺失体在哺乳动物CHO细胞中的启动子活性比较后推测:在RRD3中存在两个真核生物启动子的调控元件,一个对动物细胞的启动子起正调控,但对植物细胞中的启动子起负调控作用;另一个调控元件仅对动物细胞的启动子起负调控,而对植物细胞启动子无影响。此外在RRD3序列中至少存在一个与TATA盒相关的真核启动子核心元件,但在动物和植物细胞中的调控方式不同。
The moderate repeat RRD3 and its deletion mutant were cloned into the plant promoter vector and transformed into rice callus by Agrobacterium tumefaciens. The promoter activity in rice callus was detected by GUS histochemical method . The results showed that the full-length RRD3,410bp and 150b deletion bodies had strong promoter activity, while the 700bp and 120bp deletion bodies had only weak promoter activity. By comparing with the activity of promoter of RRD3 series deletion in mammalian CHO cells, it is speculated that there are regulatory elements of two eukaryotic promoters in RRD3, one is positive regulator of animal cell promoter, In which the promoter plays a negative regulatory role; the other regulatory element negatively regulates the promoter of animal cells only, but has no effect on plant cell promoters. In addition there is at least one eukaryotic promoter element associated with the TATA box in the RRD3 sequence, but in different ways in animal and plant cells.