Abstract
ABSTRACT: Objective To optimize the condition for transfection of HEK293 cells with exogenous gene in mediation of lipofectamineTM2000. Methods The hTERT-E1A-EGFP was inserted into the shuttle vector pEntry to construct pAd-hTERT-E1A-EGFP vector. pAd--hTERT-E1A-EGFP was then digested by PacI and transfected into the HEK293 cell line by Optimization method. Packaging recombinant adenovirus, cDNA was obtained using reverse transcription after total RNA was extracted. Results: Our results demonstrated that the adenovirus was successfully packaged by Optimization method. Conclusion The condition for transfection of HEK293 cells in mediation of lipofectamineTM 2000 was optimized. Compared with the traditional packaging recombinant adenovirus in Lipofectamine TM 2000, optimization method of packing is more effective and convenient, it not only improving the packing efficiency, shortening packaging time, reducing the possibility of cells pollution, but also laing a solid foundation for subsequent adenovirus gene therapy research.
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