NKX3.1基因上游一个30bp负调控区结合蛋白的初步鉴定

Identification of binding protein to a 30 bp-negative regulation region upstream of NKX 3.1 gene

目的:鉴定NKX3.1基因上游30bp负调控区的结合蛋白。方法:人工合成30bp负调控序列,用末端转移酶对其进行地高辛标记;提取细胞核蛋白,采用电泳迁移率变动分析(EMSA)方法,检测细胞核提取液中与30bp负调控序列特异结合的核蛋白。结果:在LNCaP细胞及其他不同肿瘤细胞系核提取液中,检测到与30bp负调控序列特异结合的核蛋白,但在不同的细胞系中有不同种类的结合蛋白。结论:NKX3.1基因上游的30bp负调控机制在不同的细胞中普遍存在,不同的细胞核中有不同的结合蛋白,可能涉及不同的调控机理。

Objective: To identify a binding protein to a 30 bp-negative regulation region upstream of NKX 3.1 gene. Methods: The 30 bp-sequence was synthesized and labeled with digoxigenin by terminal transferase. The nucleic extracts were isolated and bound to the labeled probe of 30bp-sequence. Electrophoresis mobility shift assay (EMSA) was used to identify the binding proteins to the 30 bp-sequence. Results: The proteins that bound specifically to 30 bp-sequence were found in the nucleic extracts of LNCaP cell line and other tested tumor cell lines. However, the binding proteins to 30 bp-sequence were different in various tested cell lines. Conclusions: The event of 30 bp-sequence-involved negative regulation for NKX 3.1 gene is common in different cell lines. Different binding proteins and mechanisms may be involved in the regulation of NKX 3.1 gene in different cells.