病程相关基因启动子片段与GUS的融合基因在拟南芥中的表达

PR1是拟南芥(Arabidopsisis thaliana L.)系统获得抗性的一个标志基因.利用PCR技术,从拟南芥中扩增并克隆了PR1基因的启动子片段.将该启动子片段与GUS报告基因拼接,构建成含有PR1-GUS融合基因的重组表达质粒.经根癌农杆菌介导转化,得到了转基因的拟南芥植株.用已知的系统获得抗性激活剂处理转基因植物,检测到GUS活性.因此,这一转基因体系可以作为一种简便、灵敏的实验体系以筛选激活植物系统获得抗性的化合物.

Expression of the Pathogenesis-related Gene Promoter-GUS Reporter Fusion in Arabidopsis

Pathogesis-related gene 1 (PR1) is a valid marker gene for systemic acquired resistance in Arabidopsis thaliana L. Using PCR amplification, we cloned the upstream regulatory region of PR1 gene from Arabidopsis Col-0. The fragment was then fused to the reporter gene encoding b-glucuronidase (GUS) and introduced into Arabidopsis plant by Agrobacterium-mediated gene transfer. PCR analysis and Southern blot hybridization of total DNA extracted from transgenic plants verified that the fusion gene had been integrated into the Arabidopsis genome. GUS activity can be induced by chemical treatment in transgenic Arabidopsis. The response was monitored by histochemical staining of GUS activity in situ, and fluorimetric assay in tissue extracts. Transgenic plants containing PR1-GUS or other defense gene promoter-reporter gene fusions may therefore provide specific assay systems for screening potential activators of systemic acquired resistance.