The Protein Expression and Libraries Facility (Protein Expression) provides Cancer Center members efficient and expert technical assistance in recombinant DMA plasmid engineering, 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. The Facility has 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. The proteins produced by the facility must be of high quality and purity in order to be used by Cancer Center scientists to achieve a wide range of experimental objectives, such as characterization of enzymatic activities, crystallization for structural analysis, characterization of structure-function relationships of protein:protein, protein:nucleic acid, and protein:small molecule interactions;development of assays for small molecule high throughput screening;and immunization of mice to generate custom antibodies utilizing the Hybridoma Facility. The facility will continue to expand it's repertoire of vectors available for protein expression, included high titer retroviral vectors, acquire RNAi libraries, and implement a new assay development service for highthroughput screening of small molecule libraries. The facility 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 expression plasmid construction and large-volume protein expression services, including quality assurance and control procedures to ensure efficient, consistent production and purification of high quality recombinant proteins. The Facility also maximizes biosafety by confining retrovirus (e.g. lentivirus) production to a centralized biosafety level 2 (BSL2) unit, which prevents aerosolization of viruses from contaminating incubators and parental cultures of cell lines, and improves quality control in the production of virus stocks to be used in gain- and loss-of-function experiments in vitro and in vivo. Currently the Facility is being expanded to accommodate the storage and handling of viral libraries, additional purification instruments, and the rapid assay systems to meet Cancer Center needs.

Public Health Relevance

Recombinant DMA technology has provided the unique opportunity to produce otherwise rare proteins derived from recombinant genes. The availability of these proteins has enabled many types of experiments that would have otherwise been impossible. Also, the facility will provide and manage libraries of shRNA genes in lenteviral vectors that can be used to manipulate the expression of genes for cancer research.

National Institute of Health (NIH)
National Cancer Institute (NCI)
Center Core Grants (P30)
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Subcommittee G - Education (NCI)
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Wistar Institute
United States
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Qin, Jie; Rajaratnam, Rajathees; Feng, Li et al. (2015) Development of organometallic S6K1 inhibitors. J Med Chem 58:305-14
Tomescu, Costin; Seaton, Kelly E; Smith, Peter et al. (2015) Innate activation of MDC and NK cells in high-risk HIV-1-exposed seronegative IV-drug users who share needles when compared with low-risk nonsharing IV-drug user controls. J Acquir Immune Defic Syndr 68:264-73
Gekonge, Bethsebah; Bardin, Matthew C; Montaner, Luis J (2015) Short communication: Nitazoxanide inhibits HIV viral replication in monocyte-derived macrophages. AIDS Res Hum Retroviruses 31:237-41
Webster, Marie R; Xu, Mai; Kinzler, Kathryn A et al. (2015) Wnt5A promotes an adaptive, senescent-like stress response, while continuing to drive invasion in melanoma cells. Pigment Cell Melanoma Res 28:184-95
Zhang, Xuhui; Akech, Jacqueline; Browne, Gillian et al. (2015) Runx2-Smad signaling impacts the progression of tumor-induced bone disease. Int J Cancer 136:1321-32
Kung, Che-Pei; Khaku, Sakina; Jennis, Matthew et al. (2015) Identification of TRIML2, a novel p53 target, that enhances p53 SUMOylation and regulates the transactivation of proapoptotic genes. Mol Cancer Res 13:250-62
Wolf, Amaya I; Strauman, Maura C; Mozdzanowska, Krystyna et al. (2014) Pneumolysin expression by streptococcus pneumoniae protects colonized mice from influenza virus-induced disease. Virology 462-463:254-65
Gumireddy, Kiranmai; Li, Anping; Kossenkov, Andrew V et al. (2014) ID1 promotes breast cancer metastasis by S100A9 regulation. Mol Cancer Res 12:1334-43
Budina-Kolomets, Anna; Balaburski, Gregor M; Bondar, Anastasia et al. (2014) Comparison of the activity of three different HSP70 inhibitors on apoptosis, cell cycle arrest, autophagy inhibition, and HSP90 inhibition. Cancer Biol Ther 15:194-9
Newhart, Alyshia; Janicki, Susan M (2014) Seeing is believing: visualizing transcriptional dynamics in single cells. J Cell Physiol 229:259-65

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