Simultaneous insertion of two expression cassettes into adenovirus vectors.
We have designed AdenoQuick, a fast and versatile method to construct first-generation adenoviral vectors that contain one or two transgenes in the E1 and/or the E3 region. The method is based on the reconstitution of the entire genome of the desired recombinant virus in E. coli and the subsequent transfection of the DNA in a helper cell line. Since the construction of large adenoviral plasmids is generally difficult and therefore rebuffing for inexperienced researchers, we have optimized the cloning strategy by using bacterial positive-selection markers and a set of specific restriction enzymes that allow for directional cloning. The system is 99% efficient and allows one to insert simultaneously two expression cassettes into the E1 and E3 regions of the adenovirus genome.
O.D. 260 Inc., Mountain States Medical Research Institute, and Department of Veterans Affairs Medical Center, Boise, ID, USA. xdanthin@od260.com
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Danthinne
D000256:Adenoviridae / Q000235:genetics*; D005822:Genetic Vectors; D011995:Recombination, Genetic; D019076:Transgenes