摘要
从对照和用ＤＥＨＰ处理的大鼠肝脏提取核蛋白，以含酰基ＣｏＡ氧化酶（ＡＯＸ）基因表达调控部位的ＤＮＡ片段和该基因的不同蛋白结合位点的ＤＮＡ片段作为核蛋白结合反应的探针，通过凝胶电泳迁移率改变实验和Ｓｏｕｔｈｗｅｓｔｅｒｎ印迹分析检查了ＤＥＨＰ对ＡＯＸ基因反式作用因子的影响。结果表明，降血脂药物ＤＥＨＰ可显著增加ＡＯＸ基因反式作用因子的含量和（或）与基因的结合活性，在转录水平上促进基因的表达。
The liver nuclear proteins were extracted from the control and DEHP-treated wistar rats. Utilizing the radioactive DNA fragments containing upstream regulatory elements of AOX gene and protein-binding site A and B DNA fragments as probes, we have investigated the effect of hypolipidemic drug(DEHP) on the trans-acting factor(s) for AOX gene by electrophoretic band-shift experiment and Southwestern blot analysis. The results suggest that rat liver nuclear extracts contain a protein of 100kD which exhibits specific binding activity for the binding site B ,and that the 100kD protein was markedly increased in the liver of DEHP-treated rats. These data provide evidence for the presence of the trans-acting factor(s)that can be induced by DEHP in rat liver nuclei, which may be involved in the molecular events responsible for transcriptional regulation of AOX gene.
关键词
反式作用因子
降血脂药
酰基
辅酶A氧化酶
基因
AOX gene
Trans-acting factor
DEHP
Electrophoretic band-shift
Southwestern blot
