西瓜wml1 5'端启动子区域对番茄的遗传转化及其转录调控功能研究

根据wml1 5’端启动子区域内部的限制性酶切位点,分离得到长度分别为1573bp、1197bp、896bp、795bp的片段,并与GUS基因融合构成转录融合体。用农杆菌介导法将这些片段转入番茄中,对转基因植株进行GUS活性分析,发现1573bp、1197bp、896bp的片段都能诱导GUS在授粉后15天、30天、45天的番茄果实中表达,且表达强度随果实发育而增强,而在叶片、茎、根中未检测到GUS基因表达。而795bp的片段转化的植株中则未检测到GUS基因表达。推定857bp至957bp之间的序列中包含了启动子行使正常功能必需的元件。

TRANSFORMATION OF wml1 5' PROMOTER REGION INTO TOMATO PLANTS AND STUDIES ON ITS TRANSCRIPTIONAL REGULATION ROLE

According to its restriction sites, fragments of 1573bp, 1197bp, 896bp and 795bp were obtained from the 5' promoter region of wml 1 and fused with the coding sequence of the GUS gene. Constructs containing these fragments were introduced into tomato plants via Agrobacterium -mediated transformation. Histochemical assay of GUS expression in transgenic tomato plants revealed that fragments of 1573bp, 1197bp, 896bp were able to direct GUS expression in fruits of 15, 30, 45 days after anthesis with the expression level of GUS increasing with fruit development, but not in leaves, stems and roots. While no GUS expression was observed in tomato plants transformed by construct containing fragment of 795bp. It was determined that the region from 857bp to 957bp contains the elements necessary for directing fruit-specific expression.