毛细管电泳技术分析PDGF-B基因启动子与蛋白质的复合物

摘要

With the use of 1．0%T,0%C linear polyacrylamide as sieving matrix, 0．25×TBE(Tris 89 mmol/L,boric acid 89 mmol/L, EDTA 2 mmol/L) as running buffer and 15 ℃ as column temperature, the human PDGF-B promoter binding nuclear protein can be determined within 50 min with good resolution． The results proved that there are two proteins having strong ability binding human PDGF-B promoter, which similar to that in slab gel electrophoresis． This technique can provide one of the rapid and accurate separation methods in the studying of the formation and repression behavior of DNA binding protein based on PDGF gene as the target．%利用1．0%T,0%C的线性聚丙烯酰胺(即丙烯酰胺和双丙烯酰胺的总质量分数是1．0%，交联度为0%)作为筛分介质，对人体血小板长生因子(PDGF)-B基因启动子与核蛋白形成的复合物进行了分析。结果显示主要有两种核蛋白与PDGF-B基因启动子具有较强的结合能力，能形成DNA蛋白质复合物。所得结论与传统凝胶电泳相似。该方法具有较高的分离度和良好的重现性，整个分析可在50 min内完成。该方法不失为一种基于PDGF为研究目标、用于PDGF与蛋白质复合物形成及抑制行为分析的快速、准确的实用方法。