A cis-acting sequence, located at -450 in the promoter of the human interferon-inducible gene 6-16, binds constitutively to a nuclear protein and decreases the expression of a reporter interferon-inducible promoter.

Lymphokine research Pub Date : 1990-01-01
Y Chernajovsky, H M Kirby-Sanders
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Abstract

An interferon-inducible synthetic promoter was constructed with oligonucleotides which correspond to two regions of the promoter of the human interferon-inducible gene 6-16 that interact in-vitro with nuclear proteins. Firstly, we cloned a direct repeat sequence that was necessary for interferon regulation and that interacted with a 55 kilodalton nuclear protein. 5' to the direct repeat we introduced a 45 bp long oligonucleotide located at -450 in the native 6-16 promoter that interacted in-vitro with an 80 kilodalton nuclear protein. Expression after transfection of these plasmids showed that the direct repeat is necessary for interferon-inducible control and the -450 oligonucleotide by itself has no effect on transcriptional activity while in conjunction with the direct repeat it decreased the basal and interferon-inducible transcriptional activity of the reporter promoter in human cells.

位于人类干扰素诱导基因6-16启动子-450位的顺式作用序列,组成性地与核蛋白结合,并降低报告干扰素诱导启动子的表达。
利用与人干扰素诱导基因6-16启动子的两个区域相对应的寡核苷酸构建了干扰素诱导合成启动子,这些区域在体外与核蛋白相互作用。首先,我们克隆了干扰素调控所必需的直接重复序列,该序列与55千道尔顿的核蛋白相互作用。在直接重复序列的5'处,我们引入了一个45 bp长的寡核苷酸,位于天然6-16启动子的-450位置,它在体外与一个80千道尔顿的核蛋白相互作用。转染这些质粒后的表达表明,直接重复是干扰素诱导控制所必需的,-450寡核苷酸本身对转录活性没有影响,而与直接重复一起,它降低了人细胞中报告子启动子的基础转录活性和干扰素诱导的转录活性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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