CONSTRUCTION OF ADENOVIRUS VECTORS

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Adenoviral vector-mediated gene transfer is particularly attractive for cells that are refractory to standard methods of transfection, such as calcium phosphate precipitation, liposomes or electroporation. This includes primary cell cultures and other highly differentiated cells, including non-dividing cells. For the same reasons, adenoviral vectors are the method of choice for in situ gene transfer. In addition, adenoviral vectors may be used to express several exogenous genes in a cell, either simultaneously or at defined intervals. Levels of gene expression can be controlled by the dose of virus used. The purpose of this project is to construct replication-deficient vectors for in vitro use. These vectors will direct the expression of a wide range of proteins that are of interest to investigators in the NIDR Division of Intramural Research. The vectors will be used to study signal transduction, extracellular matrix and salivary gland physiology. Specifically, the first goal of the contract will be to grow and purify plasmids containing adenovirus sequences. The second will be to recombine plasmids to generate adenovirus vectors. The third goal of the project will be to amplify isolated adenovirus plaques and to purify them into virus stocks. These reagents will be unique and not available from commercial sources.