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编码16kDa DNA结合及凋亡诱导蛋白C1D的基因的启动子。

Promoter of the gene encoding the 16 kDa DNA-binding and apoptosis-inducing C1D protein.

作者信息

Rothbarth K, Hunziker A, Stammer H, Werner D

机构信息

Division of Biochemistry of the Cell (B0300), German Cancer Research Center, Im Neuenheimer Feld 280, D-69120 Heidelberg, Germany.

出版信息

Biochim Biophys Acta. 2001 Apr 16;1518(3):271-5. doi: 10.1016/s0167-4781(01)00198-1.

Abstract

The 5' region of the gene encoding the human 16 kDa DNA-binding and apoptosis-inducing C1D protein was analysed for promoter activity. Sections of this region were cloned into a promoterless vector containing the enhanced green fluorescent protein (EGFP) as reporter gene. Expressed EGFP was estimated in transfected cells by quantitative fluorescence microscopy. The sequence between mRNA positions ATG -868 and ATG -12 results in relatively highest EGFP expression in transiently transfected human and murine cells. The upstream segment immediately adjacent to the 5' end of the most active fragment was identified as an inverted LINE-1 repeat element. Transient transfection experiments point to the presence of cis-acting repressing sequences on this LINE-1 element which reduce the transcriptional activity of the basal C1D promoter in human and murine cells by more than 95%. This result supports previous evidence suggesting that LINE-1 sequences may function as regulatory elements to control the expression of nearby genes.

摘要

对编码人类16kDa DNA结合及凋亡诱导蛋白C1D的基因的5'区域进行了启动子活性分析。该区域的片段被克隆到一个无启动子载体中,该载体含有增强型绿色荧光蛋白(EGFP)作为报告基因。通过定量荧光显微镜对转染细胞中表达的EGFP进行评估。mRNA位置ATG -868和ATG -12之间的序列在瞬时转染的人类和鼠类细胞中导致相对最高的EGFP表达。紧邻最活跃片段5'端的上游片段被鉴定为一个反向LINE-1重复元件。瞬时转染实验表明,该LINE-1元件上存在顺式作用的抑制序列,这些序列可使人类和鼠类细胞中基础C1D启动子的转录活性降低95%以上。这一结果支持了先前的证据,表明LINE-1序列可能作为调控元件来控制附近基因的表达。

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