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.
Danthinne
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O.D. 260 Inc., Mountain States Medical Research Institute, and Department of Veterans Affairs Medical Center, Boise, ID, USA. xdanthin@od260.com
D000256:Adenoviridae / Q000235:genetics*; D005822:Genetic Vectors; D011995:Recombination, Genetic; D019076:Transgenes