Gidekel M, Jimenez B, Herrera-Estrella L
Departamento de Ingeniería Genética de Plantas, Centro de Investigación y de Estudios Avanzados del I.P.N., México.
Gene. 1996 May 8;170(2):201-6. doi: 10.1016/0378-1119(95)00837-3.
Genomic and cDNA clones coding for elongation factor-1 beta (eEF-1 beta) from Arabidopsis thaliana (At) were isolated and characterized. eEF-1 beta was found to be encoded by a single-copy At gene. Chimeric genes fusing the promoter and the 5' untranslated region of the At eEF-1 beta gene to the gus reporter gene were constructed and used to study the expression of this gene in transgenic tobacco plants. Interestingly, it was found that the first intron of this gene is required for high levels of expression. Experiments using chimeric promoters showed that an enhancer-like element is present in the first intron of At eEF-1 beta. Gel-shift assays were used to demonstrate that this intron is specifically bound by putative transcription factors present in nuclear protein extracts.
从拟南芥(At)中分离并鉴定了编码延伸因子-1β(eEF-1β)的基因组和cDNA克隆。发现eEF-1β由一个单拷贝的At基因编码。构建了将At eEF-1β基因的启动子和5'非翻译区与gus报告基因融合的嵌合基因,并用于研究该基因在转基因烟草植物中的表达。有趣的是,发现该基因的第一个内含子是高水平表达所必需的。使用嵌合启动子的实验表明,At eEF-1β的第一个内含子中存在一个增强子样元件。凝胶迁移试验用于证明该内含子与核蛋白提取物中存在的假定转录因子特异性结合。