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小鼠干扰素-β1启动子的分离与功能特性分析

Isolation and functional characterization of the murine interferon-beta 1 promoter.

作者信息

Dirks W, Mittnacht S, Rentrop M, Hauser H

机构信息

Genetics and Cell Biology Section, GBF-Gesellschaft für Biotechnologische Forschung mbH., Braunschweig, FRG.

出版信息

J Interferon Res. 1989 Feb;9(1):125-33. doi: 10.1089/jir.1989.9.125.

Abstract

A murine cosmid clone harboring the single-copy interferon-beta 1 (IFN-beta 1) gene and extended flanking sequences was isolated. The functional IFN-beta 1 promoter is contained within a 170-bp DNA fragment located 5' of the coding sequence. This was shown by fusion of this fragment to a heterologous reporter gene and transient as well as stable expression in mouse L and monkey CV-1 cells. With the help of these functional assays, it could be demonstrated that the 5'-flanking sequences are the target for the typical regulatory action of common type I IFN activators. DNA sequencing reveals a considerable homology to the human IFN-beta 1 promoter within the 280 upstream base pairs. The homology is particularly pronounced within the DNA region containing the virus responsive element (VRE). This phenomenon may explain the similarity of both genes in the mode of regulation. The mouse promoter fragment compared with the human equivalent was shown to be several times more efficient in transcriptional activation in murine and primate cells.

摘要

分离出了一个携带单拷贝干扰素-β1(IFN-β1)基因及延伸侧翼序列的小鼠黏粒克隆。功能性IFN-β1启动子包含在编码序列5'端的一个170bp DNA片段内。将该片段与异源报告基因融合,并在小鼠L细胞和猴CV-1细胞中进行瞬时及稳定表达,证实了这一点。借助这些功能分析,可以证明5'侧翼序列是常见I型干扰素激活剂典型调节作用的靶点。DNA测序显示,在280个上游碱基对内,与人类IFN-β1启动子有相当高的同源性。在包含病毒反应元件(VRE)的DNA区域内,这种同源性尤为明显。这一现象可能解释了两个基因在调节模式上的相似性。与人类等效片段相比,小鼠启动子片段在小鼠和灵长类细胞的转录激活中效率要高几倍。

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