心肌转录因子和锌指转录因子在先天性心脏病中的相互作用

发布时间：2018-04-16 15:20

本文选题：心脏发育相关基因 + 基因突变　；参考：《中国临床药理学杂志》2017年18期

【摘要】：目的研究心脏发育相关基因心肌转录因子NKX2.5和锌指转录因子GATA4在先天性心脏病(CHD)的相互作用关系。方法构建基因NKX2.5和GATA4真核表达质粒及脑钠肽启动子荧光素酶报告基因表达质粒,真核表达质粒经转染后,利用免疫共沉淀技术证明基因NKX2.5和GATA4两者是否有相互作用,通过荧光素酶分析技术分析基因NKX2.5和GATA4对脑钠肽启动子序是否存在协同激活作用。结果 p CMV-HA-NKX2.5与Pre-immune作用后蛋白表达量0,与Anti-Myc作用后蛋白表达量为3.63±1.23,与Input作用后蛋白表达量为2.95±1.05,NKX2.5可将GATA4沉淀下来;p CMV-Myc-GATA4与Pre-immune作用后蛋白表达量0,与Anti-Myc作用后蛋白表达量为5.63±2.06,与Input作用后蛋白表达量为5.75±2.86,GATA4可将NKX2.5沉淀下来;脑钠肽(BNP)启动子荧光表达质粒与两者共转染比分别单独转染NKX2.5和GATA4对脑钠肽启动子的激活作用更强。结论 NKX2.5和GATA4之间具有相互作用,在心脏发育过程中起着重要的协同作用。
[Abstract]:Objective to study the interaction between cardiac development-related gene NKX2.5 and zinc finger transcription factor (GATA4) in congenital heart disease (CHD).Methods the eukaryotic expression plasmids of gene NKX2.5 and GATA4 and the plasmids expressing luciferase reporter genes of brain natriuretic peptide promoter were constructed. After transfection of eukaryotic expression plasmids, the interaction between NKX2.5 and GATA4 was proved by immunoprecipitation.Luciferase analysis was used to determine whether the gene NKX2.5 and GATA4 had synergistic effects on brain natriuretic peptide promoter sequence.Results the protein expression of p CMV-HA-NKX2.5 and Pre-immune was 0, that of Anti-Myc was 3.63 卤1.23, that of Input was 2.95 卤1.05NKX2.5, that of GATA4 precipitated by p CMV-Myc-GATA4 and Pre-immune was 0, and that of Anti-Myc was 0.After treatment with Input, the protein expression was 5.75 卤2.86 and GATA4 could precipitate NKX2.5.The fluorescent expression plasmid of brain natriuretic peptide (BNP) promoter and co-transfection of NKX2.5 and GATA4 had stronger effect on the activation of brain natriuretic peptide promoter.Conclusion there is an interaction between NKX2.5 and GATA4, which plays an important synergistic role in cardiac development.
【作者单位】：广州医科大学附属第三医院心内科;
【基金】：广东省医学科研基金面上基金资助项目(B2015134)
【分类号】：R541.1

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