Cleavage of hairpin ribozyme to target a transcribed VEGF RNA in vitro  

作　　者：严瑞兰 金明 钱新华 惠宏襄 辛晓燕 王健 王检波 

出　　处：《Journal of Medical Colleges of PLA(China)》2001年第2期102-106,共5页中国人民解放军军医大学学报（英文版）

基　　金：This work is supported by program of National Natural Science Foundation of China(39700147)

摘　　要：Objective:To constructan expressionvectorbearinghairpinribozyme(Rz)geneagainstvascularendo-thelialgrowthfactor(VEGF)andto assaythecleavageactivityof theribozymein vitro.Methods :Anti-VEGFhair-pinRz genewassynthesizedwhileVEGFgenewasclonedfromhumanplacenta,andtheywerebothinsertedinto theeukaryoticexpressionvectorpcDNA3.ThenpcDNA3-RzandthepcDNA-VEGFweretranscriptedin vitro and cleavageactivityof theRz wasmeasured.Results:Thefragmentof theRz genewasconfirmedby thedigestionof pcDNA3-RzwithEcoR I andBam HI.Theexpressionproductof theRzgenehashighercleavageactivity.Conclusion:WehavesuccessfullyconstructedpcDNA3-Rzexpressionvectorandlaida basisforfurtherstudyof gene therapy.Objective:To constructan expressionvectorbearinghairpinribozyme(Rz)geneagainstvascularendo-thelialgrowthfactor(VEGF)andto assaythecleavageactivityof theribozymein vitro.Methods :Anti-VEGFhair-pinRz genewassynthesizedwhileVEGFgenewasclonedfromhumanplacenta,andtheywerebothinsertedinto theeukaryoticexpressionvectorpcDNA3.ThenpcDNA3-RzandthepcDNA-VEGFweretranscriptedin vitro and cleavageactivityof theRz wasmeasured.Results:Thefragmentof theRz genewasconfirmedby thedigestionof pcDNA3-RzwithEcoR I andBam HI.Theexpressionproductof theRzgenehashighercleavageactivity.Conclusion:WehavesuccessfullyconstructedpcDNA3-Rzexpressionvectorandlaida basisforfurtherstudyof gene therapy.

关 键 词：vascularendothelialgrowthfactor hairpinribozyme GENETHERAPY 

分 类 号：R346[医药卫生—基础医学]