The objective of this proposal is to form a core facility for AIDS-related molecular biology with emphasis on expression and production of viral or cellular proteins for use in AIDS research. We have extensive expertise in application of several recombinant systems for expression of proteins in bacteria, established culture cell lines, and insect cell systems, which have been previously used by our laboratories to produce viral proteins of HIV-1, and HIV-2 and SIV, cellular transcription factors, growth factors, and cytokines. In addition, we have a great deal of experience in the use of various procedures for purifying individual protein compartments from culture media for cell lysates. Particular importance will be laced on use of purification procedures that will yield antigens in their native antigenic configuration. The expressed proteins will be provided to CFAR investigators for ongoing projects in vaccine development, viral pathogenesis, structure-function studies, and other research programs. In addition to providing a facility for protein production and purification, the facility will provide training to CFAR investigators in basic molecular biology techniques related to gene expression. These include training of new investigators in basic molecular biology techniques related to gene expression. These include training of new investigations in basic procedures such as plasmid preparation, transfection, and protein purification. Finally, this Core Facility will provide a repository for expression vectors and cells which will be useful for expression of cloned genes, as well as a repository of plasmids containing genes of HIV, SIV, viral receptors and co-receptors, and other related genes of interest. The overall objective is therefore to facilitate interdisciplinary research by providing essential expertise in molecular biology with particular emphasis on expression of genes of interest for AIDS research.
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