中文摘要：

目的：构建细胞分裂周期蛋白2（Cdc2）基因启动子荧光素酶报告基因载体pGL3-Cdc2-promoter，并检测其转录活性。方法根据UCSC软件查找的人基因组DNA的Cdc2启动子序列并设计两端引物，扩增人基因组DNA中的Cdc2启动子。用限制性内切酶SacⅠ和XhoⅠ双酶切质粒pGL3-Basic和Cdc2启动子后，将Cdc2基因启动子插入到pGL3-Basic报告基因载体上。重组质粒命名为pGL3-Cdc2-promoter。将其与内参质粒pRL-SV40瞬时共转染骨肉瘤细胞U2OS，检测双荧光素酶活性。结果成功构建Cdc2基因启动子荧光素酶报告基因载体pGL3-Cdc2-promoter，质粒酶切及测序结果完全正确。瞬时共转染pGL3-Cdc2-promoter/pRL-SV40组荧光素酶活性为1.5915±0.1998，高于转染pGL3-Basic/pRL-SV40组的荧光素酶活性值0.0499±0.0104。结论 pGL3-Cdc2-promot-er在骨髓瘤细胞U2OS细胞中能被转录激活，为进一步将其用于抗肿瘤药物的筛选与评价奠定了基础。

英文摘要：

Objective To construct the luciferase reporter gene vector of cell division cycle 2 （Cdc2） gene promoter and determine its transcriptional activity. Methods Primers were designed based on human Cdc2 promoter sequence from UCSC software. Then Cdc2 promoter from human genome DNA was replicated. After pGL3-Basic vector and Cdc2 promoter were digested with restriction enzymes SacⅠand XhoⅠseparately, Cdc2 promoter was inserted into pGL3-Basic vector. The recombinant plasmid named pGL3-Cdc2-promoter was transiently co-transfected into U2OS cells with control vector pRL-SV40, and then the activity of dual luciferase was detected. Results pGL3-Cdc2-promoter was constructed successfully. The restriction analysis and sequencing proved the entirely correct sequencing results. The luciferase activity was higher in pGL3-Cdc2-promoter/pRL-SV40 group than that of pGL3-Basic/pRL-SV40 group （1.591 5±0.199 8 vs 0.049 9±0.010 4）. Conclusion pGL3-Cdc2-promoter can be transcribed and activated in U2OS cells. This study provided an important basis for screening and evaluation of anticancer drugs.