An innovative system is proposed for the utilization of the adenoviral terminal protein to facilitate the construction of adenoviral vectors used for gene transfer, and in particular gene therapy. So far, adenoviral vectors are best generated using bacterial plasmids, because they eliminate potential viral contaminations. However, plasmid-derived adenoviral DNAs are poorly infectious, and the recovery of the virus sometimes can take several weeks. There is ample evidence in literature, and our preliminary data have confirmed, that the presence of the terminal protein at both ends of the adenoviral DNA increases its infectivity by two or three orders of magnitude. Our goal is to increase the infectivity of adenoviral DNAs prepared from plasmids, by using the terminal protein. We propose 1) a method for purifying the terminal protein and, b) a method to bind the purified protein to the ends of adenoviral DNAs derived from the plasmids. We brought together a strong team and the resources necessary to demonstrate our concept. We expect this development will provide a more expedient and thus less expensive method for the construction of first-generation, second-generation and gutless adenoviral vectors, than currently available.
NOT AVAILABLE
