The Protein Expression and Proteomics (PEP) Core received supplemental funding for one year startingJune 1, 2006, to provide an unmet need for our community of investigators. The Protein Expression portionof the proposed pilot Core, under the direction of Dr. John Elder, will provide the appropriate vectors andcell lines needed to facilitate expression cloning of investigator-targeted proteins. The Proteomics portion ofthe core, under the direction of Dr. Bruce Torbett, will provide protein analysis services for CFARinvestigators. These services include protein extraction from viruses, cells, and tissues; protein resolutionand quantification using 1- and 2-dimensional (D) electrophoresis and imaging systems; proteinidentification using mass spectroscopy; and analyses of proteins using Luminex xMAP multiplexdetection technology. The unique aspect of the PEP Core is the focus on providing services to investigatorsrequiring expression of proteins from HIV-1 and other viral genes and protein analysis of HIV-1 and othervirally infected samples.The Protein Expression portion of the Core will provide the appropriate vectors and cell lines needed tofacilitate expression cloning of investigator-targeted proteins, using bacterial, insect cell, or mammalian cellvector systems, as deemed appropriate for the particular application. The Elder laboratory has compiled anextensive panel of vectors for each expression system and has extensive experience in protein expressionand purification for ligand binding studies, enzymology, and crystallographic analyses. The laboratory isequipped with incubators required to grow cells at the required temperatures and also centrifuges, FPLC,and HPLC systems to aid in protein purification where necessary. The Core will aid the investigator invector selection and amplification and cloning of targeted genes. In addition, the Core will help investigatorsin optimizing protein expression and selection of purification schemes most appropriate for eachapplication.The Proteomics portion of the Core will provide all facets of protein resolution from visualization toidentification. CFAR members will have access to the NCI-funded Proteomics Core housed within theDepartmental of Molecular and Experimental Medicine at The Scripps Research Institute. Currently theProteomics Core has available 1- and 2-D gel electrophoresis protein separation instrumentation, IEFZoom fractionators for solution phase IEF, a state-of-the-art 2D Proteomic Imaging System with a Bio-RadVersaDoc 1000 imager and imaging software. All imaging software can be utilized free of charge onresident computers housed within the Core. Technical advice is available from the Core technicians, andwhen needed, Core technicians provide demonstrations and seminars on analysis and imaging software. Inaddition, Core technicians will provide advice to investigators for optimizing protein extraction proceduresand selection of purification schemes most appropriate for each application.The Protein Expression and Proteomics Core will provide informational seminars/sessions on the Coreservices each year and 'new technology' seminars/sessions throughout the funding period.
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