Buchel A S, Molenkamp R, Bol J F, Linthorst H J
Institute of Molecular Plant Sciences, Gorlaeus Laboratories, Leiden University, Netherlands.
Plant Mol Biol. 1996 Feb;30(3):493-504. doi: 10.1007/BF00049327.
The 900 bp promoter region of the tobacco PR-1a gene was divided into eight fragments using PCR. The fragments were tested for their ability to bind to nuclear factors isolated from tobacco leaf. Band shift assays demonstrated that all but one of the fragments specifically interacted with nuclear proteins. From competition experiments it was determined that the same nuclear factors bind various promoter fragments with different affinity. Moreover, efficient competition with a synthetic tetramer of box II of the rbcS promoter indicated that GT-1-like nuclear factors are involved in these interactions. Furthermore, in comparison to extracts from untreated plants, nuclear protein preparations from tobacco mosaic virus-infected tobacco showed a reduced GT-1 binding activity. These results will be discussed in relation to induced PR-1a gene expression.
利用聚合酶链式反应(PCR)将烟草PR-1a基因900 bp的启动子区域分为八个片段。检测这些片段与从烟草叶片中分离出的核因子的结合能力。凝胶迁移实验表明,除一个片段外,其他所有片段都能与核蛋白特异性相互作用。通过竞争实验确定,相同的核因子以不同亲和力结合各种启动子片段。此外,与rbcS启动子II号盒的合成四聚体的有效竞争表明,GT-1样核因子参与了这些相互作用。此外,与未处理植物的提取物相比,感染烟草花叶病毒的烟草的核蛋白制剂显示出降低的GT-1结合活性。将结合诱导型PR-1a基因表达对这些结果进行讨论。