Marcotte W R, Russell S H, Quatrano R S
Molecular Biology, Central Research and Development, E.I. du Pont de Nemours and Co., Inc., Wilmington, Delaware 19880-0402.
Plant Cell. 1989 Oct;1(10):969-76. doi: 10.1105/tpc.1.10.969.
We demonstrate that a chimeric gene containing the beta-glucuronidase (GUS) reporter gene linked to a 646-base pair 5' fragment (-554 to +92) from the abscisic acid (ABA)-regulated Em gene from wheat is correctly expressed in transgenic tobacco. We observe high activity only in embryos of mature seeds, and immature seeds cultured on ABA show enhanced expression. Using a rice transient assay, we identify a 260-base pair fragment (-168 to +92) that accounts for the ABA-specific 15-fold to 20-fold increase in GUS expression. A 50-base pair sequence (-152 to -103) fused 5' in either orientation to a truncated cauliflower mosaic virus promoter (35S) increases GUS activity threefold in the presence of ABA. Insertion of the Em 5'-untranslated region (+6 to +86) between the 35S promoter and the ATG of GUS results in a 10-fold increase in GUS activity in the absence of ABA. These results suggest the following two functional fragments of the Em 5' region: an ABA response element from -152 to -103 and an element between +6 and +86 that quantitatively increases the ABA response.
我们证明,一个嵌合基因在转基因烟草中能够正确表达,该嵌合基因包含与来自小麦脱落酸(ABA)调控的Em基因的646个碱基对的5'片段(-554至+92)相连的β-葡萄糖醛酸酶(GUS)报告基因。我们仅在成熟种子的胚中观察到高活性,并且在添加ABA培养的未成熟种子中表达增强。使用水稻瞬时分析,我们鉴定出一个260个碱基对的片段(-168至+92),该片段导致GUS表达中ABA特异性的15至20倍增加。一个50个碱基对的序列(-152至-103)以任一方向与截短的花椰菜花叶病毒启动子(35S)5'端融合,在ABA存在下使GUS活性增加三倍。在35S启动子和GUS的ATG之间插入Em 5'非翻译区(+6至+86),在没有ABA的情况下使GUS活性增加10倍。这些结果表明Em 5'区域的以下两个功能片段:一个从-152至-103的ABA响应元件和一个在+6和+86之间的元件,该元件定量增加ABA响应。