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The pattern of high mobility group proteins 1 and 2 (HMG1,2) interaction with the 5'-flanking sequence of the human β-globin gene has been analyzed by scanning tunnelling microscopy (STM). A 200 bp negative regulatory region in the 5'-flanking sequence of the human β-globin gene can be folded by HMG proteins 1 and 2 into a circular structure (diameter 70±6) with a linear tail which seems to be a left-handed double helix structure. 相似文献
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Our previous studies have identified that there are at least three regulatory regions (two negative regions and one positive region) in the 5'-flanking sequence of human β-globin gene (-610 to +1 bp). The binding of HMG proteins to both negative regulatory regions was examined by the gel mobility shift and DNase I protection assays.In gel mobility shift assay,we observed that HMG proteins 1 and 2 could bind to both negative regulatory regions (NCR1 and NCR2).Using the gel shift competition assay,we identified that the binding proteins between the two regions are different from each other.DNase I protection analysis shows that HMG proteins 1 and 2 only bind to one site (between-560 and-533 bp) in NCR1.However,two protected regions can be detected in NCR2, one between-272 and-252 bp relative to the cap site, the other between-306 and-329 bp.We also observed that HMG proteins 14 and 17 could not bind to both negative regions, so it seems that HMG proteins 1 and 2 may play an important role in the regulati 相似文献
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人体β-珠蛋白基因的5′-旁侧DNA片段(-610bp→+1bp)内的两个负调控区域与HMG蛋白质(High Mobility Group Proteins)结合状况已用近代分子生物学的技术(足印分析法(DNase Ⅰ protection Assay)和凝胶电泳阻抑分析法(Gel MobilityShift Assays)加以分析。用凝胶电泳阻抑分析方法证明了HMG蛋白质(1+2)与两个负调控区域有不同程度的结合,同时也观察到这两个负调控区域的结合蛋白是各异的。为了进一步检测HMG蛋白质(1+2)与DNA序列的精确结合位点,我们又采用了足印分析的方法,观察到在第一个负调控区内HMG蛋白质(1+2)有一个专一的结合位点(-560bp→-533bp)。在第二个负调控区内,则可以检测到有两个结合位点(-272bp→-252bp和-306bp→-329bp)。另外,我们还证实了HMG蛋白质(14+17)与这两个负调控区都不能结合。上述实验结果提示HMG蛋白质(1+2)在β-珠蛋白基因表达中起着积极的调控作用。 相似文献
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