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Adenoviruses Expressing Cytosine Deaminase Enerated by Adenovirus Plasmid Homologous Recombinant in Bacteria and Observation of Its Tumor-Killing Effect in vitro
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Abstract:
Objective: To generate an adenovirus, Ad-CD, expressing green fluorescence protein and bacterial cytosine deaminase, and use it for tumor suicide gene therapy. Methods: CD gene was cloned into pAdTrack-CMV, pAdTrack-CMV-CD and pAdEasy-1 were homologous recominated in bacteria. 293 cells were transfected by the newly-recombinated plasmid pAd-CD and generated adenoviruses. LoVo cells were infected with the adenovirus, and the tumor-killing effect after 5-FC was administered.Results: pAdTrack-CMV-CD and the newly-recombinated plasmid pAd-CD were tested by restriction endonuclease digestions. Followed by GFP, Ad-CD were produced in 293 and LoVo cells infected by Ad-CD were killed after 5-FC was administered.Conclusion: Adenovirus plasmid homologous recombinant in bacteria is a simple method for adenovirus generation. In vitro adenoviral carried CD gene is efficient for tumor cell suicide gene therapy and has a bystand-effect, which is not depended on cell direct contact
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Clc Number:
R737.31
