White R A, Brookman J J, Gould A P, Meadows L A, Shashidhara L S, Strutt D I, Weaver T A
Department of Anatomy, University of Cambridge, UK.
J Cell Sci Suppl. 1992;16:53-60. doi: 10.1242/jcs.1992.supplement_16.7.
We have used a chromatin immunopurification approach to identify target genes regulated by the homeotic gene Ultrabithorax. A monoclonal antibody against the Ultrabithorax gene product is used to immunopurify in vivo Ultrabithorax protein binding sites in embryonic chromatin. The procedure gives an enrichment of sequences with matches to a consensus homeodomain binding site. In one case we have shown that an immunopurified sequence lies within a 4 kb fragment that acts in vivo as a homeotic response element. We anticipate that this approach will enable us to identify further targets, allowing the analysis of their regulation and function. The chromatin immunopurification strategy may be of general application for the identification of direct in vivo targets of DNA-binding proteins.
我们采用了染色质免疫纯化方法来鉴定由同源异型基因超双胸调控的靶基因。一种针对超双胸基因产物的单克隆抗体被用于在胚胎染色质中免疫纯化体内超双胸蛋白结合位点。该程序能富集与共有同源结构域结合位点相匹配的序列。在一个案例中，我们已表明一个免疫纯化的序列位于一个4 kb的片段内，该片段在体内作为一个同源异型反应元件发挥作用。我们预计这种方法将使我们能够鉴定更多的靶标，从而对它们的调控和功能进行分析。染色质免疫纯化策略可能在鉴定DNA结合蛋白的直接体内靶标方面具有广泛应用。
