Curie C, Liboz T, Bardet C, Gander E, Médale C, Axelos M, Lescure B
Laboratoire de Biologie Moléculaire des Relations Plantes-Microorganismes, CNRS-INRA, Castanet Tolosan, France.
Nucleic Acids Res. 1991 Mar 25;19(6):1305-10. doi: 10.1093/nar/19.6.1305.
In A. thaliana the translation elongation factor EF-1 alpha is encoded by a small multigenic family of four members (A1-A4). The A1 gene promoter has been dissected and examined in a transient expression system using the GUS reporter gene. Deletion analysis has shown that several elements are involved in the activation process. One cis-acting domain, the TEF 1 box, has been accurately mapped 100 bp upstream of the transcription initiation site. This domain is the target for trans-acting factors identified in nuclear extracts prepared from A. thaliana. Homologies are found between the TEF 1 box and sequences present at the same location within the A2, A3 and A4 promoters. This observation, together with those obtained from gel retardation assays performed using DNA fragments from the A4 promoter, suggest that the activation process mediated by the TEF 1 element is conserved among the A. thaliana EF-1 alpha genes. Analysis of nearly full length cDNA clones has shown that in addition to a single intron located within the coding region, the A1 gene contains a second intron located within the 5' non coding region. Such an intron is also present within the A2, A3 and A4 genes. This 5' intervening sequence appears to be essential to obtain a maximum GUS activity driven by the A1 gene promoter.
在拟南芥中,翻译延伸因子EF-1α由一个包含四个成员(A1 - A4)的小多基因家族编码。已使用GUS报告基因在瞬时表达系统中对A1基因启动子进行了解剖和检测。缺失分析表明,有几个元件参与激活过程。一个顺式作用结构域,即TEF 1框,已被精确地定位在转录起始位点上游100 bp处。该结构域是从拟南芥制备的核提取物中鉴定出的反式作用因子的作用靶点。在TEF 1框与A2、A3和A4启动子中相同位置存在的序列之间发现了同源性。这一观察结果,连同使用来自A4启动子的DNA片段进行凝胶阻滞分析所获得的结果,表明由TEF 1元件介导的激活过程在拟南芥EF-1α基因中是保守的。对几乎全长的cDNA克隆的分析表明,除了位于编码区内的一个单一内含子外,A1基因在5'非编码区内还含有第二个内含子。这样的内含子也存在于A2、A3和A4基因中。这个位于5'端的间隔序列似乎对于获得由A1基因启动子驱动的最大GUS活性至关重要。