Core B - David C. Schultz, Ph. D. The Protein and Vector Production Core will be responsible for time efficient and expert technical assistance in the management of project resources, protein expression in bacteria and baculovirus infected insect cells, purification of recombinant proteins to homogeneity, and production of high-titer stocks of retroviruses (e.g. lentiviruses) for delivery of shRNA and cDNAs to mammalian cells. Expertise in all aspects of vector technology for protein expression, baculovirus generation, recombinant protein expression, and affinity and conventional chromatography approaches to protein purification, and production of infectious lenti- /retroviruses will be provided to all projects on a continual basis for the lifetime of grant support. The Core personnel are highly trained technical experts in all areas of the proposed support services to be provided to the individual projects. The centralization and standardization of these practices under the direction of very experienced staff allows for high-throughput, large-volume, protein expression services, including quality assurance and control procedures to ensure efficient, consistent production and purification of high quality recombinant proteins. Furthermore, confinement of retrovirus (e.g. lentivirus) production to a centralized biosafety level 2 (BSL2) unit provides increased biosafety, prevention of aerosolized viruses from contaminating incubators and parental cultures of cell lines, and quality control in the production of virus stocks to be used in experiments. In addition, the collection of projects in this proposal is expected to generate unique research resources that will be required for programmatic interactions. The Core will function as a repository for collection, initial processing, storage, maintenance, and distribution of newly developed materials. In this manner, resources can be reliably tracked to ensure quality control, adherence to guidelines for resource sharing, and consistency in usage within programmatic interactions and the greater scientific community.
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