Recombinant Virus and Vector Core

摘要(见说明)：重组工程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)在巴氏杆菌KSHV中产生定点突变，以及4)大规模生产重组和野生型KSHV。这些目标将通过一个集中的实验室来实现，提供一套标准化的服务，包括疱疹病毒、分子工程和基于重组的突变(即条件敲除的Kan-SACB、Kan-rpsL、Gal-K和loxP-Kan)的最先进方法，生产用于cDNA和shRNA的重组Lenti病毒表达载体，以及从转基因293细胞(重组病毒)、具有特定突变的GFP标记病毒或BCBL1细胞(野生型病毒)中生产KSHV。由于预计这一方案项目将产生独特的研究资源，核心将负责储存重组病毒、杆菌、重组工程和Lenti病毒生产的试剂，以及管理用户可以通过基于网络的浏览器访问和搜索的适当数据库。在赠款的有效期内，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.