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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 basisforfurtherstudyofgene therapy.