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大鼠生长抑素基因细胞特异性表达相关启动子序列的鉴定

Identification of the promoter sequences involved in the cell specific expression of the rat somatostatin gene.

作者信息

Andrisani O M, Hayes T E, Roos B, Dixon J E

出版信息

Nucleic Acids Res. 1987 Jul 24;15(14):5715-28. doi: 10.1093/nar/15.14.5715.

Abstract

DNA sequences containing the 5' flanking region of the rat somatostatin gene were linked to the coding sequence of the bacterial chloramphenicol acetyl transferase gene. This recombinant plasmid is active in expressing CAT activity in the neuronally derived, somatostatin producing CA-77 cell line. Deletion analyses of the somatostatin promoter show that the sequences proximal to position -60, relative to the cap site are required for expression of this promoter. A 4 base pair deletion of residues -46 through -43 within the somatostatin promoter results in a down mutation in vivo suggesting the existence of an element critical for the expression of the promoter in CA-77 cells. In addition, the somatostatin recombinant and its 5' deletion constructs preferentially express CAT activity in CA-77 cells, whereas only basal level of expression is observed in HeLa, BSC40, and RIN-5F cell lines, pointing to the cell specific nature of this promoter.

摘要

含有大鼠生长抑素基因5'侧翼区的DNA序列与细菌氯霉素乙酰转移酶基因的编码序列相连。这种重组质粒在神经元来源的、产生生长抑素的CA - 77细胞系中能够有效表达氯霉素乙酰转移酶(CAT)活性。对生长抑素启动子的缺失分析表明,相对于帽位点,靠近 - 60位置的序列是该启动子表达所必需的。生长抑素启动子内 - 46至 - 43位残基的4个碱基对缺失在体内导致了一个负向突变,这表明存在一个对CA - 77细胞中启动子表达至关重要的元件。此外,生长抑素重组体及其5'缺失构建体在CA - 77细胞中优先表达CAT活性,而在HeLa、BSC40和RIN - 5F细胞系中仅观察到基础水平的表达,这表明该启动子具有细胞特异性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63a7/306017/22f487d97dbc/nar00258-0219-a.jpg

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