Chernajovsky Y, Kirby-Sanders H M
Department of Immunology, University of Texas M.D. Anderson Cancer Center, Houston.
Lymphokine Res. 1990 Summer;9(2):199-212.
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.
用与人类干扰素诱导基因6 - 16启动子的两个区域相对应的寡核苷酸构建了一个干扰素诱导型合成启动子,这两个区域在体外与核蛋白相互作用。首先,我们克隆了一个干扰素调节所必需的直接重复序列,该序列与一个55千道尔顿的核蛋白相互作用。在直接重复序列的5'端,我们引入了一个位于天然6 - 16启动子中- 450位置的45 bp长的寡核苷酸,该寡核苷酸在体外与一个80千道尔顿的核蛋白相互作用。这些质粒转染后的表达表明,直接重复序列对于干扰素诱导控制是必需的,而- 450寡核苷酸本身对转录活性没有影响,但与直接重复序列结合时,它会降低人类细胞中报告基因启动子的基础转录活性和干扰素诱导转录活性。