Institut für Genbiologische Forschung Berlin, Ihnestrasse 63, 1000 Berlin 33, Federal Republic of Germany.
Proc Natl Acad Sci U S A. 1987 Nov;84(22):7943-7. doi: 10.1073/pnas.84.22.7943.
ST-LS1 is a light-inducible, single-copy gene from potato that is expressed only in photosynthetic tissues. Various sequences derived from the 5'-upstream region of this gene were fused to the coding region of the chloramphenicol acetyltransferase (CAT) gene and to the gene 7 termination region of the transfer DNA (T-DNA) from the Agrobacterium Ti plasmid pTiACH5 and transferred to tobacco using Ti-plasmid vectors. After regeneration of whole plants, tissues were assayed for the expression of the CAT gene. Sequences derived from the 5'-upstream region of the ST-LS1 gene comprising positions -334 to +11 were sufficient to confer a leaf/stem-specific as well as a light-inducible expression of the CAT gene. Destruction of chloroplasts by treatment with the herbicide norfluorazon and subsequent exposure to light drastically reduced the expression of the CAT gene indicating that this upstream sequence most likely interacts with a chloroplast-dependent signal. When sequences from position -98 to position +675 were fused to a truncated inactive fragment of the cauliflower mosaic virus 35S promoter in a head-to-head manner, the corresponding chimeric genes were again expressed in photosynthetic tissues only, indicating that these sequences have enhancer-like properties.
ST-LS1 是一种来自马铃薯的光诱导、单拷贝基因,仅在光合组织中表达。该基因 5'上游区域的各种序列与氯霉素乙酰转移酶 (CAT) 基因的编码区域以及农杆菌 Ti 质粒 pTiACH5 的转移 DNA (T-DNA) 的基因 7 终止区域融合,并使用 Ti 质粒载体转移到烟草中。在再生整个植株后,对 CAT 基因的表达进行组织分析。来自 ST-LS1 基因 5'上游区域的序列,包含位置 -334 到 +11,足以赋予 CAT 基因叶片/茎特异性以及光诱导表达。用除草剂 norfluorazon 处理破坏叶绿体,随后暴露在光线下,大大降低了 CAT 基因的表达,表明该上游序列很可能与叶绿体依赖的信号相互作用。当从位置 -98 到位置 +675 的序列以头对头的方式融合到花椰菜花叶病毒 35S 启动子的截短非活性片段上时,相应的嵌合基因仅在光合组织中表达,表明这些序列具有增强子样特性。