基于mPGES-1表达调控的药物筛选模型的建立  

Establishment of a Drug Screening Model Based on Regulating Sequences of mPGES-1

作　　者：郭雷[1] 王淑军[1] 龚邦强[2] 

机构地区：[1]淮海工学院海洋学院,连云港222005 [2]华东理工大学生物工程学院,上海200237

出　　处：《天然产物研究与开发》2008年第B05期1-4,共4页Natural Product Research and Development

基　　金：淮海工学院引进人才科研项目（KQ07026）

摘　　要：聚合酶链式反应扩增mPGES-1上游调控序列并插入到已预先在pGL3-Basic载体的Sal Ⅰ位点插入真核筛选基因新霉素抗性基因（neomycin）形成的质粒pGL3B—neo中，构建重组报告基因载体pGL3B-neo／mPGES-1，转染人肺癌细胞A549，通过G418筛选，建立了稳定的mPGES-1表达下调剂筛选细胞株SPGES1。通过检测荧光素酶报告基因表达水平的变化筛选人mPGES．1表达下调剂，并优化了其筛选条件，优化的条件是每孔细胞接种数目为1×10^4-3×10^4个，溶剂DMSO终浓度为1％，底物用量为20μL。利用该模型可有效地进行人mPGES-1表达下调剂的筛选。The upstream regulatory sequences of mPGES-1 were obtained by polymerase chain reaction （PCR） and cloned into pGL3B-neo vector （the pGL3B-neo vector had been constructed by cloning the neomycin resistance gene into the Sal I site of the pGL3-Basie vector） ,the recombinant reporter gene vector pGL3B-neo/mPGES-1 was transfected into A549 cells to make stable cell line. The assay conditions were optimized and the optimal conditions were that the inoc- ulation cell numbers are 1 ×10^4-3× 10^4 ,the final concentration of DMSO is 1% and the substrate volume is 20μL in each well. This screening model can be applied to identify down-regulators Of mPGES-1.

关 键 词：膜结合型前列腺素E2合酶-1 炎症 前列腺素E2 报告基因 筛选模型 

分 类 号：R965.1[医药卫生—药理学]