Theill L E, Wiborg O, Vuust J
Department of Molecular Biology and Plant Physiology, University of Aarhus, Denmark.
Mol Cell Biol. 1987 Dec;7(12):4329-36. doi: 10.1128/mcb.7.12.4329-4336.1987.
Fragments of 5'-flanking and noncoding exon I sequences of the human gastrin gene were analyzed in transient expression assays after transfection of a variety of cell lines with the pSVCAT vector system. In the presence of the simian virus 40 (SV40) enhancer, the gastrin gene fragment from nucleotides -250 to +57, relative to the cap site, was as efficient a promoter as the SV40 early promoter itself. In the absence of the SV40 enhancer, gastrin gene 5'-flanking sequences had no promoter activity except in the murine neuroblastoma cell line N18TG2. In this cell line, the fragment from -1300 to +57 stimulated transcription as actively as the SV40 early promoter with its enhancer. This cell-specific gastrin gene promoter activity was in accordance with the finding that gastrin is synthesized in certain neuronal cells. Promoter activity declined with decreasing distance from the 5' end to the cap site and disappeared after removal of the gastrin gene TATA box. In vector constructions containing short vector-linker sequences homologous to a functionally important region of the SV40 enhancer, the gastrin gene fragment from -17 to +57 showed considerable promoter activity, exclusively in N18TG2. It is concluded that the truncated gastrin gene promoter plus the first exon contains a cell-specific element that may act in collaboration with upstream elements to facilitate the accumulation of transcripts.
采用pSVCAT载体系统转染多种细胞系后,在瞬时表达分析中对人胃泌素基因5'-侧翼和非编码外显子I序列的片段进行了分析。在猿猴病毒40(SV40)增强子存在的情况下,相对于帽位点,从核苷酸-250至+57的胃泌素基因片段作为启动子的效率与SV40早期启动子本身相同。在没有SV40增强子的情况下,胃泌素基因5'-侧翼序列除在小鼠神经母细胞瘤细胞系N18TG2中外没有启动子活性。在该细胞系中,从-1300至+57的片段刺激转录的活性与带有增强子的SV40早期启动子一样。这种细胞特异性的胃泌素基因启动子活性与胃泌素在某些神经元细胞中合成的发现一致。启动子活性随着从5'端到帽位点距离的减小而下降,在去除胃泌素基因TATA框后消失。在含有与SV40增强子功能重要区域同源的短载体接头序列的载体构建体中,从-17至+57的胃泌素基因片段仅在N18TG2中显示出相当的启动子活性。得出的结论是,截短的胃泌素基因启动子加上第一个外显子包含一个细胞特异性元件,该元件可能与上游元件协同作用以促进转录本的积累。