CORE--GENE TRANSFER

基因转移核心的功能是为复制缺陷重组腺病毒的构建、筛选、大规模纯化和滴定提供便利。重组腺病毒为成人心肌细胞高效基因转移和直接改变表型提供了可靠的方法。具体地说，该核心将1)维持人293内皮肾细胞系，2)扩增和维持腺病毒穿梭质粒和腺病毒骨架质粒的纯化库，3)将重组腺病毒DNA导入293 HEK细胞，4)在293 HEK细胞中繁殖重组腺病毒，5)对重组腺病毒进行空斑纯化并进行大规模纯化，以及6)在293 HEK细胞中通过空斑分析测定纯化的腺病毒滴度。该核心还将负责重组腺病毒的长期存储，并将协助个人研究人员在成人心肌细胞中执行基因转移方案。一名全职研究专家已经接受了运行这一核心所涉及的所有程序和技术的培训，并将接受核心主任的监督。

The function of the Gene Transfer Core is to provide a facility for the construction, screening, large-scale purification, and titrating of replication-defective, recombinant adenoviruses. Recombinant adenoviruses provide a reliable method for high efficiency gene transfer into adult cardiocytes and for directly altering the phenotype. Specifically, the core will 1) maintain lines of human 293 endothelial kidney cells [HEK], 2) amplify and maintain purified stocks of adenovirus shuttle plasmids and adenovirus backbone plasmids, 3) transfect recombinant adenoviral DNA into 293 HEK cells, 4) propagate recombinant adenoviruses in 293 HEK cells, 5) plaque-purify recombinant adenoviruses and perform large-scale purifications, and 6) titer purified adenoviruses by plaque assays in 293 HEK cells. The core will also be responsible for long-term storage of recombinant adenoviruses and will assist individual investigators in carrying out gene transfer protocols in adult cardiocytes. A full-time research specialist has been trained in all of the procedures and techniques involved in running this core, and will be supervised by the core director.