PLAGL2对SP-C基因表达调控的作用靶点研究

Study the interaction site of PLAGL2 modulating surfactant protein-C gene expression

目的研究PLAGL2对SP-C基因表达调控的作用靶点。方法定点诱变技术分别突变掉SP-C基因启动子上PLAGL2的结合位点以及PLAGL2的第6、7位锌指结构域序列并获得相应的突变体;凝胶电泳迁移实验研究PLAGL2及其突变体分别与SP-C基因启动子及其突变体之间的相互结合作用。结果PLAGL2能与同位素标记的SP-C基因启动子片段相结合形成蛋白-DNA复合物,而两者的突变体却不能与相应的蛋白或DNA片段发生相互作用。结论PLAGL2通过其第6?7位锌指结构与SP-C基因启动子上PLAGL2的核心和簇结合位点序列相互结合从而实现对SP-C基因的表达调控。

Objective： To study the interaction site of PLAGL2 modulating surfactantprotein - C gene expression Methods ： Site - directed mutagenesis was performed to get the SP - C gene promoter mutant （ SP - CP - mComclu） and PLAGL2 mutants （ PLAGL2 - mF6, PLAGL2 - mF7） ; EMSA was performed to identified the ability and specffity of interaction between PLAGL2 or it＇s mutant and SP- C gene promoter DNA fragment or it＇s mutant. Results： EMSA and competitive EMSA result showed that in vitro translated PLAGL2 protein can bind to SP - C gene promoter probe to form the protein - DNA retarded complex, but their mutants cant. Conclusion： PLAGL can bind to it＇s core and cluster binding sequence on SP - C gene promoter probe to form the protein - DNA retarded complex by it＇s zinc finger 6 and 7.