The Recombineering and Virus Production core will be responsible for 1) recombinant virus design consultation, 2) vector and viral reagent repository; 3) generation of site-directed mutations in Bacmid KSHV, and 4) large scale production of recombinant and wild-type KSHV. These goals will be accomplished by a centralized laboratory, providing a set of standardized services, including state ofthe art methods in herpes virus, molecular engineering and recombination-based mutagenesis (i.e. Kan-SacB, Kan-rpsL, Gal-K, and LoxP-kan for conditional knock-outs), production of recombinant Lenti-virus expression vectors for cDNA and shRNA, and production of KSHV from transfected 293 cells (recombinant viruses), GFP tagged viruses with specific, mutations, or BCBL1 cells (wild-type virus). Since it is expected that this program project will generate unique resources for research, the core will be responsible for storage of recombinant viruses, bacmids, reagents for recombineering and Lenti-virus production, as well as management of adequate databases that users can access and search via a web-based browser. Expertise in all aspects of protein and virus production will be provided to the projects for the lifetime of grant support.
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