2012•Unpublished venueRequires access
Construction of Recombinant Adenovirus-endostatin Vector and Detection of Its Expression
Long Hong-qing
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Abstract
Objective To construct a recombinant adenovirus vector expressing human endostatin and to detect the expression of Ad-Endostatin in HEK293 cells.Methods Endostatin gene was cloned into plasmid pShuttle2 vector and pAdeno-X vector.Recombinant pAd-Endostatin was tansformated into HEK293 cells and Ad-Endostatin viruses were obtained,purified and titered.Endostatin expression was analyzed by Western Blot.Results pAd-Endostatin was successfully constructed and verified by PCR and sequencing.Ad-Endostatin was reproduced and endostatin was highly expressed in HEK293 cells.8 days after transfection,CPE could be observed in HEK293 cells by microscope.Conclusion The recombinant plamid pAd-Endostatin with endostatin gene was constructed successfully.After HEK293 cells are infected by Ad-Endo recombinant viruses,Endostatin can be expressed in the cells.