Pruzina S, Hanscombe O, Whyatt D, Grosveld F, Philipsen S
National Institute for Medical Research, Mill Hill, London, UK.
Nucleic Acids Res. 1991 Apr 11;19(7):1413-9. doi: 10.1093/nar/19.7.1413.
The Locus Control Region (LCR) of the human beta globin gene domain is defined by four erythroid-specific DNasel hypersensitive sites (HSS) located upstream of this multigene cluster. The LCR confers copy number dependent high levels of erythroid specific expression to a linked transgene, independent of the site of integration. To assess the role of the individual hypersensitive sites of the LCR, we have localized HSS4 to a 280bp fragment that is functional both in murine erythroleukaemia (MEL) cells and in transgenic mice. This fragment coincides with the major area of hypersensitivity 'in vivo' and contains a number of DNasel footprints. Bandshift analysis shows that these footprints correspond to binding sites for the erythroid specific proteins GATA1 and NF-E2 and a number of ubiquitous proteins, including jun/fos, Sp1 and TEF2.
人类β珠蛋白基因结构域的位点控制区(LCR)由位于该多基因簇上游的四个红系特异性脱氧核糖核酸酶超敏位点(HSS)所界定。LCR赋予与它相连的转基因以拷贝数依赖的高水平红系特异性表达,且与整合位点无关。为评估LCR中各个超敏位点的作用,我们已将HSS4定位到一个280bp的片段上,该片段在小鼠红白血病(MEL)细胞和转基因小鼠中均具有功能。此片段与“体内”超敏的主要区域相符,并包含多个脱氧核糖核酸酶足迹。凝胶迁移分析表明,这些足迹对应于红系特异性蛋白GATA1和NF-E2以及一些普遍存在的蛋白质(包括jun/fos、Sp1和TEF2)的结合位点。