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大鼠γ-晶状体蛋白基因的组织和物种特异性启动子元件

Tissue- and species-specific promoter elements of rat gamma-crystallin genes.

作者信息

Peek R, van der Logt P, Lubsen N H, Schoenmakers J G

机构信息

Department of Molecular Biology, University of Nijmegen, The Netherlands.

出版信息

Nucleic Acids Res. 1990 Mar 11;18(5):1189-97. doi: 10.1093/nar/18.5.1189.

Abstract

The 5' flanking regions of the six rat gamma-crystallin genes (gamma A-gamma F) are all capable of conferring lens-specific expression to the bacterial chloramphenicol acetyl transferase (CAT) reporter gene in either transdifferentiating chicken neural retina cells or mouse lens epithelial cells. Deletion mapping of the most active gamma-crystallin promoter region, the gamma D region, showed that at least three elements are required for maximal expression in mouse lens epithelial cells: element(s) located between -200 and -106, a conserved CG rich region around position -75, and a CG stretch around -15. The region between -200 and -106 was dispensable in transdifferentiating chicken neural retina cells, which instead required the region between -106 and -78. The maximal activity of the gamma E and gamma F promoters was also dependent upon the integrity of the conserved CG region located around -75. A synthetic oligonucleotide containing this sequence was capable of lens-specific enhancement of the activity of the tk promoter in transdifferentiating chicken neural retina cells but not in mouse lens epithelial cells. Our results further show that this region may contain a silencer element, active in non-lens tissues, as well.

摘要

六个大鼠γ-晶状体蛋白基因(γA - γF)的5'侧翼区域,在分化中的鸡神经视网膜细胞或小鼠晶状体上皮细胞中,均能够使细菌氯霉素乙酰转移酶(CAT)报告基因实现晶状体特异性表达。对活性最强的γ-晶状体蛋白启动子区域,即γD区域进行缺失定位分析表明,在小鼠晶状体上皮细胞中实现最大表达至少需要三个元件:位于-200至-106之间的元件、-75位置附近富含CG的保守区域以及-15附近的CG序列。在分化中的鸡神经视网膜细胞中,-200至-106之间的区域并非必需,取而代之的是需要-106至-78之间的区域。γE和γF启动子的最大活性也取决于-75位置附近保守CG区域的完整性。包含该序列的合成寡核苷酸能够在分化中的鸡神经视网膜细胞中对tk启动子的活性进行晶状体特异性增强,但在小鼠晶状体上皮细胞中则不能。我们的结果进一步表明,该区域可能还包含一个在非晶状体组织中也有活性的沉默子元件。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/19fb/330434/37182c0753f2/nar00189-0096-a.jpg

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