A more effective and easier method for constructing recombinant adenovirus based on AdEasy system

  Objective   To Obtain a more effective and easier method for constructing recombinant adenovirus by simplifying the protocol of the AdEasy system.

  Methods   The replication-defective adenovirus type 5 backbone plasmid pAdEasy-1 was transformed into competent E. coli BJ5183 by electroporation and the E. coli BJ5183/p was obtained.Then E.coli BJ5183/p competent cells were prepared by CaCl ₂ technique and the shuttle plasmid carrying green fluorescent protein(GFP)was transformed into E. coli BJ5183.The confirmed recombinant was linearlized with Pac I and transfected into 293 cell line.

  Results   The CPE of 293 cells were observed and the green fluorescence was seen under fluorescent microscope.

  Conclusion   A more effective and easier method for constructing recombinant adenovirus based on AdEasy was obtained successfully.