Li Yun-Feng, Zhu Rui, Xie Long-Xu, Xu Pei-Lin
The Key Laboratory of Gene Engineering of the Education Ministry, Zhongshan University, Guangzhou 510275, China.
Zhi Wu Sheng Li Yu Fen Zi Sheng Wu Xue Xue Bao. 2005 Apr;31(2):143-8.
A promoter of the gene encoding beta-1, 3-glucanase isoenzyme GIII was amplified from barley genomic DNA using PCR. The GIII gene promoter, designated P(GIII), was ligated upstream of the gus report gene and pGIII-gus fusion fragment was then cloned into a binary vector pCAMBIA1300 for Agrobacterium-mediated transformation of rice (Oryza sativa L. cv. Taipei 309). PCR analyses indicated that the fusion gene was present in all T0 transgenic plants. The integration of the gene into rice genomic DNA was further confirmed by Southern blot. Histochemical staining and spectrofluorophotometric analyses of transgenic rice leaves showed that the GUS activity was increased after treatment with SA and fungal elicitor. Northern blot analysis also showed expression of such induction. Histochemical staining of T(1) rice seeds displayed marked GUS activity after induction with SA and elicitor, while no GUS activity was detected in untreated T(1) rice seeds. The results presented here strongly suggested that P(GIII) could be pathogen-inducible promoter.
利用聚合酶链式反应（PCR）从大麦基因组DNA中扩增出编码β-1,3-葡聚糖酶同工酶GIII的基因启动子。将该GIII基因启动子（命名为P(GIII)）连接到gus报告基因的上游，然后将pGIII-gus融合片段克隆到双元载体pCAMBIA1300中，用于农杆菌介导的水稻（Oryza sativa L. cv. Taipei 309）转化。PCR分析表明，融合基因存在于所有T0转基因植株中。通过Southern杂交进一步证实了该基因整合到水稻基因组DNA中。对转基因水稻叶片进行组织化学染色和荧光分光光度分析表明，用水杨酸（SA）和真菌激发子处理后，GUS活性增强。Northern杂交分析也显示了这种诱导表达。T(1)代水稻种子的组织化学染色显示，用SA和激发子诱导后，GUS活性显著，而未处理的T(1)代水稻种子未检测到GUS活性。此处给出的结果强烈表明，P(GIII)可能是一种病原体诱导型启动子。
