Construction of recombinant adenoviral vector by a simple, cheap,and high performance method

Objective To set up a basis for treatment of malignant tumor with recombinant adenovirus carrying suicide genes establish a simple cheap and high efficient method of construction of recombinant adenoviral vector. Methods Adenoviral shuttle plasmid pAdTrack-CMV was linearized by PmeⅠand then purified by gel. Mixed the purified digest and adenoviral backbone plasmid pAdEasy-1 with proper proportion and transformed them into E.coli BJ 5 183 with chemical transformation method. In BJ 5 183 homologous recombination was occurred and recombinant adenoviral plasmid rpAdEasyGFP was generated. Recombinant plasmid was then transfected into 293 cells by liposome method to package recombinant adenovirus rAdGFP. Results The rate of homologous recombination was up to 80%. Recombinant adenovirus was produced in 293 cells successfully. Conclusion AdEasy system a broad applied method presently was improved. The new method can obtain higher positive recombinant rate as well as reduce the requirement for experimental equipments.