[An easier method for generating recombinant adenovirus containing CD/TK fusion gene driven by vascular endothelial growth factor promoter].

Objective: To efficiently construct the recombinant adenovirus containing CD/TK fusion gene driven by vascular endothelial growth factor (VEGF) promoter using improved homologous recombination in bacteria.

Methods: Chemical transformation, instead of electroporation, of the plasmid pAdEasy-1 into E.coli BJ5183 strain was performed to prepare BJ5183pAdEasy-1 as the competent bacterium, in which pAdEasy-1 and pAdtrack-VEGFP-CDglyTK were recombined. Finally, the recombinant adenovirus of AdVEGFP-CDglyTK was constructed by transfecting 293 cells with linearized adenoviral genomes of pAdEasy-VEGFP-CDglyTK.

Results: This new transformation procedure generated recombinant plasmids in a yield of 60% (6/10), and the adenovirus AdVEGFP-CDglyTK was harvested 7-12 d after transfection.

Conclusion: The improved homologous recombination in bacteria is efficient, convenient and easy to be carried out.