内含子和5'非翻译区对人血小板生成素基因表达的影响  

Role of intron and 5' untranslated region in human thrombopoietin gene expression

作　　者：宁云山[1] 李妍[1] 周明乾[2] 李明[1] 曾溢滔 王小宁[2] 

机构地区：[1]第一军医大学热带病研究所,广东广州510515 [2]第一军医大学分子免疫研究所,广东广州510515 [3]上海医学遗传研究所,上海210009

出　　处：《第一军医大学学报》2004年第9期991-994,共4页Journal of First Military Medical University

基　　金：国家科委"1035"重大项目(96-901-009);广东省自然科学基金重点项目(963011)~~

摘　　要：目的观察人血小板生成素(TPO)基因组中内含子及5'非翻译区对TPO表达的影响。方法分别构建含TPO基因组中不同内含子的各类TPO 真核表达质粒,在cos-1细胞中进行了瞬间表达,并采用高灵敏定量试剂盒检测培养上清中人血小板生成素的含量。结果在转染48 h后,各类TPO重组质粒在cos-1细胞中表达顺序为:TPO intron v> TPOcDNA>ΔTPO intron I> TPO intron I> TPO gDNA。结论在TPO基因组中,最后一个内含子可显著提高TPO的表达水平,表明其可能含有特殊的增强序列;同时进一步证实,在TPO基因组中可能存在抑制TPO高水平表达的结构元件,造成TPO基因组在细胞中不表达或低水平表达。Objective To study the role of 5' untranslated region (UTR) and intron in the expression of human thrombopoietin (TPO) gene. Methods A number of expression vectors containing TPO mini-gene fused to the regulatory elements of cytomegalovirus (CMV) were constructed and transfected via lipofectin into cultured cos-1 cells for transient expression of TPO gene. The cell culture media were analyzed with highly sensitive sandwich enzyme-linked immunosorbent assay (ELISA) 48 h after the transfection. Results The expression levels of the TPO gene elements followed the order of TPO intron v> TPOcDNA>ΔTPO intron I> TPO intron I> TPO gDNA in cos-1 cells. Conclusion The last intron of TPO gene obviously enhances the expression level of TPO gene, which can be inhibited by 5`UTR of TPO gene.

关 键 词：血小板生成素 内含子 基因表达 

分 类 号：R346[医药卫生—基础医学]