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Cis-acting sequences from mouse rDNA promote plasmid DNA amplification and persistence in mouse cells: implication of HMG-I in their function

机译:小鼠rDNA的顺式作用序列促进质粒DNA在小鼠细胞中的扩增和持久性:HMG-1在其功能中的意义

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Searching for amplification promoting sequences within the murine rDNA cistrons, we isolated two elements from the nontranscribed spacer region. These 370 bp and 423 bp long cis-acting elements, refered to as muNTS1 and muNTS2, are localized 4.1 kb and 4.6 kb upstream the RNA polymerase I transcriptional start site. They contain Ca. 50 bp long AT-rich sequences that strongly interact with a protein from nuclear extracts. The protein could be purified and identified as HMG-I. A synthetic oligonucleotide encompassing the AT-rich stretch from muNTS1 is able to substitute for the muNTS elements. A similar sequence from the nontranscribed spacer of rat has previously been reported to be important for the function of the RNA polymerase I enhancer (1). Therefore the interaction of HMG I with the muNTS elements may play a role both in the stimulation of DNA amplification and transcription.
机译:搜索鼠rDNA顺反子内的扩增促进序列,我们从非转录间隔区中分离出两个元素。这些370 bp和423 bp长的顺式作用元件(称为muNTS1和muNTS2)位于RNA聚合酶I转录起始位点上游4.1 kb和4.6 kb。它们包含钙。 50 bp长的富含AT的序列,可与核提取物中的蛋白质强烈相互作用。该蛋白可以被纯化并鉴定为HMG-1。包含来自muNTS1的富含AT的延伸序列的合成寡核苷酸能够替代muNTS元件。先前已经报道过,来自大鼠非转录间隔区的相似序列对于RNA聚合酶I增强子的功能很重要(1)。因此,HMG I与muNTS元件的相互作用可能在DNA扩增和转录刺激中均起着作用。

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