拟南芥PR-1基因启动子的克隆与植物表达载体的构建

Cloning Pathogenesis-related Gene Promoter of Arabidopsis and Construction of Plant Expression Vector

根据已报道的拟南芥(Arabidopsis thaliana L.)病程相关蛋白基因(PR-1)序列设计引物,通过PCR技术从拟南芥中扩增得到水杨酸诱导表达的PR-l基因启动子片段,序列分析表明,该启动子含910bp核苷酸,与已报道的序列比较,核苷酸的同源性为99.7%。将该启动子构建到植物表达载体pBI121上,获得病程相关蛋白基因(PR-1)启动子驱动的GUS报告基因的植物表达载体pBI-pr1p,将其转入根癌农杆菌GV3101,通过农杆菌介导转化拟南芥,获得转基因拟南芥植株,为深入研究寄主-病原物相互作用的分子机理奠定基础。

One pair of primer was designed based on the pathogenesis - related protein 1 of Arabidopsis thaliana L. Using PCR amplification, the promoter of PR - 1 gene （910bp）was cloned from Arabidopsis to build inducible expression system of screening plant expression vector; by Sequence analysis, the 910 nucleotides promoter showed 99.7% homology with the reported sequence; the fragment was then instead of CaMV35S of the reporter gene encoding 3 - glucuronidase （GUS） ; so a new plant expression vector named pBI - prpl in which the reporter gene GUS is droved by promoter of pathogenesis related protein was constructed. It was transfen＇ed into Agrobacterium tumefaciens GV3101, then the transgenic Arabi-dopsis plants was obtained through Agrobacterium-mediated transformation which provided a reliable basis for the further study of host-pathogen interaction.