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Upstream sequences of rice proliferating cell nuclear antigen (PCNA) gene mediate expression of PCNA-GUS chimeric gene in meristems of transgenic tobacco plants

机译:水稻增殖细胞核抗原(PCNA)基因的上游序列介导PCNA-GUS嵌合基因在转基因烟草植物分生组织中的表达

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The transgenic tobacco plants have been generated that express the E. coli β-glucuronldase (GUS) gene under control of the promoter from the rice proliferating cell nuclear antigen (PCNA, DNA polymerase auxiliary protein) gene. GUS expression detected in situ by staining with the chromogenic substrate, 5-bromo-4-chloro-3-indolyl-β-D-glucuronide (X-Gluc), was restricted to meristems in the organs of the transgenic tobacco plants. This expression responded to the phytohormones which promote callus formation. Furthermore, in situ thymldine uptake showed that the GUS expression pattern corresponded well to the active sites of DNA synthesis. Deletion analysis of the 5′ upstream sequence confined the GUS expression pattern to a fragment extending 263 bp upstream of the transcription start site of the rice PCNA gene. Thus, we have identified this fragment as a main regulatory element of the rice PCNA gene promoter.
机译:已经产生了在来自水稻增殖细胞核抗原(PCNA,DNA聚合酶辅助蛋白)基因的启动子的控制下表达大肠杆菌β-葡萄糖醛酸糖苷酶(GUS)基因的转基因烟草植物。通过用发色底物5-溴-4-氯-3-吲哚基-β-D-葡萄糖醛酸(X-Gluc)染色原位检测到的GUS表达仅限于转基因烟草植物器官中的分生组织。该表达响应于促进愈伤组织形成的植物激素。此外,原位胸腺嘧啶的摄取表明GUS表达模式与DNA合成的活性位点很好地对应。对5'上游序列的缺失分析将GUS表达模式限制为在水稻PCNA基因的转录起始位点上游延伸263bp的片段。因此,我们已经鉴定出该片段是水稻PCNA基因启动子的主要调控元件。

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