Recombinant Virus and Vector Core

摘要（见说明书）：重组和病毒生产核心将负责1)重组病毒设计咨询，2)载体和病毒试剂库；3) Bacmid KSHV位点定向突变的产生；4)重组型和野生型KSHV的大规模生产。这些目标将通过一个集中的实验室来实现，该实验室将提供一系列标准化的服务，包括最先进的疱疹病毒、分子工程和基于重组的诱变方法（即用于条件敲除的Kan-SacB、Kan-rpsL、Gal-K和LoxP-kan），生产用于cDNA和shRNA的重组慢病毒表达载体，以及从转染的293细胞（重组病毒）、带有特定突变的GFP标记病毒中生产KSHV。或BCBL1细胞（野生型病毒）。由于预计该计划项目将产生独特的研究资源，核心将负责存储重组病毒、bacmids、重组试剂和lentiv生产，以及管理足够的数据库，用户可以通过基于web的浏览器访问和搜索。在赠款支持的整个生命周期内，将向这些项目提供蛋白质和病毒生产各方面的专门知识。

SUMMARY (See instructions): 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.