Stratagene made the production of recombinant adenovirus more efficient with the AdEasy™ adenoviral vector system. Because recombination of the gene of interest into the adenoviral backbone occurs in bacteria, the system does not require cloning into the adenoviral genome or painstaking plaque isolations. These improvements can save weeks of time (Figure 1). With the AdEasy system, the homologous recombination step is performed in the E. coli strain BJ5183. With the improved AdEasy™ XL adenoviral vector system*, the BJ5183 competent cells are pre-transformed with the adenoviral backbone, pAdEasy-1, resulting in 300% more colonies that contain the desired recombinant.

Single-Plasmid Transformation Streamlines Protocol

The AdEasy XL system includes all the components of the original system plus two improved features for faster and easier adenovirus production. The system offers BJ5183-AD-1, an electroporationcompetent strain of E. coli that has been transformed with pAdEasy-1—the adenoviral backbone. As a result, the two-plasmid transformation of pShuttle vector and pAdEasy-1 into BJ5183 has been replaced by a single-plasmid transformation of pShuttle into BJ5183-AD-1 (Figure 2). The system also comes with a specialized 293 cell line, which is optimized to adhere to plates better for improved packaging of the adenovirus.

Three-Fold Higher RecombinationEfficiency in E. coli

With the AdEasy XL adenoviral vector system, you not only save time over traditional adenoviral production systems, but with the adenoviral backbone pre-transformed into E. coli, you dramatically improve production of recombinant adenoviral vector with your desired expression cassette. Generating recombinant adenovirus vector using the original AdEasy system resulted in an 18 to 34% success rate, while the pre-transformed BJ5183-AD-1 results in consistently >94% success rate (Table 1).

BJ5183-Ad-1 Cells Available Separately

For users of the original AdEasy system, the specialty BJ5183-AD-1 electroporation-competent cells are available separately, so you can take advantage of the higher recombination frequency without having to work with a new vector system.

* See license reference 5 on page 32
** See license reference 6 on page 32