CORE--GENE TRANSFER

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Gene transfer technology either with the aid of viral vectors or in the form of naked DNA has been used extensively in recent years to elucidate mechanisms regulating vascular function and blood pressure. Preliminary data generated by project investigators with the help of the Core director indicate that this technology provides useful tools to each of the program project components. The core director collaborated with project investigators for more than ten years and recently produced and delivered recombinant retroviruses expression vectors and viruses (adenoviruses and retroviruses) containing primarily the CYP4A and HO genes for in vitro and in vivo studies proposed by the project investigators. The core will perform the following functions; 1) construct recombinant CYP4A, HO and COX-2 adenoviruses and retroviruses using commercially available vectors with and without GFP or beta-gal tagging; 2) adenoviruses and retroviruses using commercially available vectors with and without GRP or beta-gal tagging; 2) isolate and purify and propagate recombinant viral vectors; 3) determine titers and maintains stocks of recombinant viral vectors; 4) maintain stocks of the various plasmids and competent and packaging cells; 5) develop vectors carrying tissue and cell lineage specific promoters; and 6) provide the resources and expertise to each investigator for evaluating transgene expression and work along with the project investigators in validating transgene expression in vitro and in vivo using RT/PCR, Southern blot analysis and detection of GFP or beta-gal. Project investigators will be supported by the core director and the technical staff in the generation and production of molecular reagents, experimental design and data interpretation.