人PSMA基因启动子表达载体pGL3-PSMP的构建  

Construction of the expression vector pGL3-PSMP containing prostate-specific membrane atigen promoter

作　　者：康鲁东[1] 崔福爱[1] 吴伟芳[1] 胡晓燕[1] 姜安丽[1] 孔峰[1] 于清水[1] 

机构地区：[1]山东大学医学院生化与分子生物学研究所,山东济南250012

出　　处：《山东大学学报（医学版）》2005年第7期567-569,588,共4页Journal of Shandong University:Health Sciences

基　　金：山东省卫生厅基金资助课题(2003HZ042)。

摘　　要：目的:构建含人前列腺特异性膜抗原(prostatespecificmembraneantigen,PSMA)基因启动子表达载体pGL3-PSMP。方法:PCR扩增人PSMA上游1175bp启动子序列,克隆至荧光素酶表达载体pGL3-Basic,构建前列腺特异性表达载体pGL3-PSMP,重组子经双酶切、测序鉴定。将构建载体用脂质体转染体外培养的前列腺癌细胞株PC-3M,应用双荧光素酶测定系统检测荧光素酶的表达活性。结果:序列测定表明,克隆获得的1175bp的DNA序列与GenBank报道的一致,且插入方向正确。含人PSMA基因启动子的报告基因荧光素酶的表达活性显著提高,比pGL3-Basic高10倍多。结论:成功构建了含人PSMA基因启动子的荧光素酶表达载体。Objective： To construct the expression vector pGL3-PSMP containing prostate-specific membrane antigen promoter （PSMP）. Methods： PSMP was amplified with the human genome DNA by PCR, the segment was cloned into the eukaryotic expression vector pGL3-Basic. The recombinant was detected by endonuclease and sequenced. Plasmid pGL3-PSMP was transfected into PC-3M cells by lipofectamine. The activity of luciferase was detected, and the effect of the PSMA promoter was studied. Results： The sequenced segment （1 175 bp） in the recombinants was identical to that GenBank had reported and the segment was inserted in right direction. The activity of luciferase was significantly increased in the pGL3-PSMP transfected cells over 10 times than that in the pGL3-Basic＇s. Conclusion： The luciferase expression vector, pGL3-PSMP containing prostate-specific membrane antigen promoter is constructed successfully. It lays foundation for studying its tissue-specificity and target gene therapy of the prostate cancer.

关 键 词：前列腺肿瘤 前列腺特异性膜抗原 启动子 荧光素酶 

分 类 号：R737.25[医药卫生—肿瘤]