MZF-2转录因子与甲基化后的DNA结合基序结合活性检测

 目的 探讨MZF-2转录因子与甲基化后的DNA结合基序结合活性。方法 常规培养CNE-2细胞株，提取核蛋白；合成、标记和纯化MZF-2探针，其中一组用M.SssI甲基化酶处理，进行凝胶迁移分析（EMSA）。结果 EMSA图中单独加入甲基化探针组无结合条带，单独加入原样未甲基化探针组有结合条带，同时加入标记和未标记原样未甲基化探针组结合条带强度减弱。结论 甲基化可以抑制MZF-2转录因子与其DNA结合基序的结合活性。

The detection of binding between MZF-2 and its methylated DNA binding motif

Objective: To detect the binding between MZF-2 and its methylated DNA　binding motif. Metheods: CNE-2 cell lines was cultured conventionally. Nuclear extracts from cells were prepared. The MZF-2 probe was synthysized, Purified, radiolabelled with the γ-32P-labeled oligonucleotide. One of groups was treated by SssI methylase. EMSA was performed. Results: No specific band was observed in group using methylated MZF-2 probe. The specific band was observed using unmethylated MZF-2 probe. In competition experiment, unlabeled oligonucleotide DNA probe interfered with the binding between MZF-2 protein and its DNA binding motif, which make the band weak. Conclusion: Methylation may inhibit the binding between MZF-2 and its binding motif.