Yamaguchi Yube, Yoda Hiroshi, Akiyama Kenji, Sano Hiroshi
Research and Education Center for Genetic Information, Nara Institute of Science and Technology, 630-0192 Nara, Japan.
Planta. 2003 Dec;218(2):249-53. doi: 10.1007/s00425-003-1108-2. Epub 2003 Sep 16.
Immediate early responsive genes were screened by the differential display method during the hypersensitive response upon tobacco mosaic virus infection of tobacco ( Nicotiana tabacum L.) plants carrying the N gene. Three hours after temperature shift from 30 degrees C to 20 degrees C, an increase in transcripts of a particular clone was observed. The cDNA encoded a polypeptide of 330 amino acids, whose topology indicated it to be a seven-transmembrane protein, designated as Nt7TM1. This was confirmed by direct observation of cultured tobacco cells expressing an Nt7TM1-green fluorescent protein fusion protein, which migrated exclusively to the plasma membrane and the endoplasmic reticulum. RNA blot hybridization analysis indicated that Nt7TM1 transcripts were not induced by salicylic or jasmonic acids, ethylene or hydrogen peroxide. The results suggested the presence of a unique system for pathogen response involving a novel seven-transmembrane protein.
利用差异显示法,对携带N基因的烟草(Nicotiana tabacum L.)植株在烟草花叶病毒感染后的过敏反应过程中的立即早期反应基因进行了筛选。在温度从30℃转变为20℃三小时后,观察到一个特定克隆的转录本有所增加。该cDNA编码一个含有330个氨基酸的多肽,其拓扑结构表明它是一种七跨膜蛋白,命名为Nt7TM1。这一点通过直接观察表达Nt7TM1-绿色荧光蛋白融合蛋白的培养烟草细胞得到了证实,该融合蛋白仅迁移至质膜和内质网。RNA印迹杂交分析表明,水杨酸、茉莉酸、乙烯或过氧化氢均不能诱导Nt7TM1转录本的产生。结果提示存在一个涉及新型七跨膜蛋白的独特病原体反应系统。