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设施。重组腺病毒提供了一种可靠的方法，可以高效地将基因转移到成人心肌细胞中，并直接改变心肌细胞的表型。具体来说，核心将1)维持人293内皮肾细胞[HEK]系，2)扩增和维持纯化的腺病毒穿梭质粒和腺病毒主干质粒，3)将重组腺病毒DNA转染293 HEK细胞，4)在293 HEK细胞中繁殖重组腺病毒，5)对重组腺病毒进行斑块纯化并进行大规模纯化，6)通过在293 HEK细胞中进行斑块测定来滴度纯化的腺病毒。该核心还将负责重组腺病毒的长期储存，并将协助个别研究人员在成人心肌细胞中进行基因转移方案。一名全职研究专家已经接受了运行该核心所涉及的所有程序和技术的培训，并将由核心主任监督。

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.