人CYP19基因启动子荧光素酶报告基因载体的构建  

Construction of Luciferase Report Gene Vector Containing Human CYP19 Promotor

作　　者：邵喜英[1] 陈占红[1] 黄健[1] 雷蕾[1] 冯建国[2] 苏丹[2] 王晓稼[1] 

机构地区：[1]浙江省肿瘤医院,浙江杭州310022 [2]浙江省肿瘤研究所,浙江杭州310022

出　　处：《肿瘤学杂志》2010年第11期873-876,共4页Journal of Chinese Oncology

基　　金：浙江省自然科学基金(Y206856);浙江省医药卫生优秀青年科技人才专项基金(2005QN002);浙江省医药卫生科学研究基金资助项目(2007B021)

摘　　要：[目的]克隆人CYP19基因启动子PⅠ.3(-345～-234bp)、PⅡ(-517～-278bp)和PⅠ.7(-299～+81bp)片段,构建启动子荧光素酶报告基因载体。[方法]采用生物合成PⅠ.3(-345～-234bp)、PⅡ(-517～-278bp)和PⅠ.7(-299～+81bp)区域基因片段,通过HindⅢ和KpnⅠ酶切连接到pGL3-Basic载体上。[结果]测序验证pGL3-Basic-PⅠ.3,pGL3-Basic-PⅡ和pGL3-Basic-PⅠ.7构建成功。[结论]pGL3-Basic-PⅠ.3,pGL3-Basic-PⅡ和pGL3-Basic-PⅠ.7荧光素酶报告基因载体的成功构建,为进一步研究CYP19基因组织特异性表达奠定基础。[Purpose] To construct luciferase report gene vector containing human CYP19 promotor PⅠ.3（-345--234bp）,PⅡ（-517--278bp）and PⅠ.7（-299-＋81bp）.[Methods] PⅠ.3（-345-234bp）,PⅡ（-517--278bp）and PⅠ.7（-299-＋81bp） were amplified by biosynthesis,and inserted into the luciferase report gene pGL3-Basic vector with restriction endonuclease HindⅢ and Kpn Ⅰ.[Results] pGL3-Basic-PⅠ.3,pGL3-Basic-PⅡ and pGL3-Basic-PⅠ.7 were confirmed by DNA sequencing.[Conclusion] The human CYP19 promotor luciferase reporter gene vectors are constructed successfully,which establishes the base for the further research of CYP19 tissue-specific expression mechanism.

关 键 词：CYP19基因 启动子 荧光素酶 

分 类 号：R730.23[医药卫生—肿瘤]