小鼠LRP5基因负调控区分析  

Analysis on the Negative Regulatory Region of Mouse LRP5 Gene

作　　者：吕萍[1] 龚瑶琴[1] 李江夏[1] 周海斌[1] 陈丙玺[1] 

机构地区：[1]山东大学医学院医学遗传学研究所实验畸形学教育部重点实验室,山东济南250012

出　　处：《药物生物技术》2007年第3期164-167,共4页Pharmaceutical Biotechnology

基　　金：教育部科学技术研究重点项目(批准号02129)

摘　　要：为研究小鼠LRP5基因-1 229 bp^-1 034 bp之间存在的负调控区,在此区域内构建了3种荧光素酶报告基因表达载体,即pGL3-1229(-1 229 bp^-914 bp),pGL3-1130(-1 130 bp^-914 bp)及pGL3-1051(-1 051 bp^-914 bp)。以PRL-TK为内参照质粒,瞬时转染COS-7,48 h后收集细胞测定荧光素酶相对表达活性。结果表明pGL3-1229质粒的相对荧光素酶活性是pGL3-1130的26%,有显著性差异。在-1 229 bp^-1 130 bp之间存在负调控元件,软件分析COUP可能是小鼠LRP5基因的负调控元件,有待进一步突变分析证实。To study the negative control elements in the region between the-1229bp and-1034bp,the region supposed to contain the negative control elements was amplified by PCR,and PCR products were cloned into the pGL3-103 vector which contained mouse LRP5 gene basal promoter and luciferase reporter gene.Three luciferase expression constructs were constructed and sequenced,which were pGL3-1229(-1229bp～-914bp),pGL3-1130(-1130bp～-914bp)and pGL3-1051(-1051bp～-914bp).Using Lipofectimine 2000 as transfection reagent,these constructs were transfected into COS-7 cell line respectively,then the relative luciferase activity was measured after 48h.The results showed that the relative luciferase activity of pGL3-1229 was 26% of pGL3-1130.The negative control element was located from-1229bp to-1130bp.Mat Inspector V2.2 software analysis showed that COUP was the good candidate for negative control element in the region,and the mutation analysis was needed for further confirmation.

关 键 词：基因LRP5 小鼠 调控序列 荧光素酶分析 

分 类 号：R394-33[医药卫生—医学遗传学]