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

作　　者：姜安丽[1] 张鹏举[1] 胡晓燕[1] 贺美兰[1] 陈蔚文[1] 孔峰[1] 张建业[1] 

机构地区：[1]山东大学医学院生物化学与分子生物学研究所,山东济南250012

出　　处：《山东大学学报（医学版）》2005年第5期375-378,共4页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金资助项目(30171026)

摘　　要：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.

关 键 词：DNA结合蛋白 负调控 电泳迁移率变动分析 同源盒基因 

分 类 号：Q756[生物学—分子生物学]