水稻重复序列RRD3缺失体介导gusA在水稻愈伤组织中的表达

Expression of gusA in Rice Calli Mediated by Deletants of Rice Repetitive DNA Sequence RRD3

摘要:

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、410 bp及150 bp缺失体具有强的启动子活性,而700 bp、120 bp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。

关键词:水稻; 植物启动子; 瞬时表达; GUS组织化学检测

Abstract:

Rice repetitive DNA sequence RRD3 and its six serial deletants were inserted into the promoter-capture vector of plants. Thus, gusA gene can be used to assess the promoter activity of inserted DNA. The promoter activities of serial deletants were detected after co-culture of rice calli with Agrobacterium LBA4404 containing different recombinant DNA by transient GUS (beta-glucuronidase) assay. The results indicated that total length of RRD3, 410 bp and 150 bp deletants had stronger promoter activities, while the promoter activities in 700 bp and 120 bp deletants were reduced. There are some positive/negative control elements in RRD3 sequence.

Key words:Rice; Plant promoter; Transient expression; GUS histochemical assay