【摘要】 目的　进一步确定ETO抑制基因转录的区域及其与组蛋白脱乙酰化酶之间的关系。方法　以位点诱变法使ETO的两个锌指结构分别突变 ,使用聚合酶链反应扩增ETO突变体 ,克隆入真核细胞表达质粒pFA CMV中。通过基因转染、报告基因转录活性分析 ,确定ETO的不同区段对基因转录调节的作用。结果　成功构建出锌指突变和不同区段的ETO表达载体 ,并发现ETO存在两个抑制转录的区域 ,分别位于羧基末端的两个锌指结构和第 2 75～ 487位氨基酸残基。结论　ETO通过 2个区域抑制转录 ,并且与组蛋白脱乙酰化酶密切相关 ,后者可能会用作急性髓系白血病M2b的治疗靶点。更多还原

【Abstract】 Objective To further verify the transcriptional repression domains in ETO and their relationship with histone deacetylase (HDAC). Methods Either of the ETO two zinc fingers was mutated respectively by site-directing mutagenesis. The truncation fragments of ETO were amplified by polymerase chain reaction (PCR) and cloned into eukaryotic expression plasmid pFA-CMV. By the means of DNA transfection and analysis of the transcription derived from the promoter of reporter gene, the transcriptional regulation domains of ETO was determined. Results The expression plasmids carrying truncated ETO and ETO with point mutation at either zinc finger were successfully constructed. Two repression domains were found within ETO, which were located at two zinc finger motifs and 275-487 amino acid residues, respectively. Conclusion The transcription repression by ETO was mediated by two separated domains and closely associated with HDAC, which may be used as therapeutic target for acute myeloid leukemia M 2b .更多还原