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目的构建携带Smac基因、人尿路斑块蛋白Ⅰb(UroplakinⅠb,UpⅠb)启动子(promoter)调控的膀胱移行上皮特异性真核表达载体。方法切除含Smac基因的真核表达载体pcDNA3.1-Smac内部的人巨细胞病毒和T7启动子序列,替换为在膀胱移行上皮特异表达的UpⅠb的启动子。构建的质粒经双酶切凝胶电泳及测序鉴定。结果成功构建携带Smac基因人UpⅠb启动子调控的膀胱移行上皮特异性真核表达载体pcDNA3-UpⅠb promoter-Smac质粒。结论新构建的载体为膀胱癌的靶向基因治疗奠定了基础。
Objective To construct bladder transitional epithelial specific eukaryotic expression vector carrying Smac gene and Uroplakin Ⅰb promoter. Methods The human cytomegalovirus and T7 promoter sequences were excised from the eukaryotic expression vector pcDNA3.1-Smac containing Smac gene and replaced with the promoter of UpⅠb specifically expressed in bladder transitional epithelium. The constructed plasmid was identified by double enzyme digestion gel electrophoresis and sequencing. Results The bladder transitional epithelial-specific eukaryotic expression vector pcDNA3-UpIb promoter-Smac plasmid carrying Smac gene human UpⅠb promoter was successfully constructed. Conclusion The newly constructed vector lays a foundation for targeted gene therapy of bladder cancer.