【摘要】 目的构建前列腺特异的PGL3-LUC表达载体,观察报告基因的表达情况以验证其组织特异性。方法经过改造的高效前列腺特异抗原(PSA)启动子全序列经酶切亚克隆入pGL3-basic荧光素酶报告基因载体,重组载体pGL3-psa转染前列腺癌细胞(Du-145)和肠癌细胞(HT-29),并应用双荧光素酶检测系统检测荧光素酶活性,排除转染过程误差。结果转染重组载体pGL3-psa诱导前列腺癌细胞荧光素酶活性明显增强,而空载体pGL3-basic和肠癌细胞的荧光素酶表达处于基础水平,比较差异有显著性意义P<0.05。结论构建的经过改造的高效前列腺特异抗原(PSA)启动子的荧光素酶报告基因载体,转染前列腺癌细胞后能够表达荧光素酶蛋白,具有组织特异性。更多还原

【Abstract】 Objective To construct hProstate-Specific Promoter Derived from PSA and PSMA Enhancers driven fluorescein luciferase reporter plasmid and evaluate specificity of the modulation of the gene expression vector.Methods The PSA promoter was subcloned into pGL3-basic luciferase reporter plasmid.and the recombinant vector pGL3.PSA was transfected into DU145 and HT29 cells.The dual luciferase activities were assayed by dual luciferase reporter(DLR)system to eliminate the error during transfection.Results Luciferase activities were increased after induced by pGL3-psa in DU145.but they remained in the basic level induced by empty vector pGL3.Basic and HT29.with significant difference between them(P<0.05).Conclusion The constructed PSA promoter driven luciferase reporter plasmid is used to transfect DU145 cells to express luciferase protein specificity.更多还原