CORE--GENE TRANSFER

基因转移中心的功能是为 构建、筛选、大规模纯化和滴定 复制缺陷型重组腺病毒。重组腺病毒 为高效地将基因转入成人提供了可靠的方法 心肌细胞和直接改变表型。具体而言是 核心将1）维持培养中的293个肾细胞，2）扩增并维持 纯化的pdeltaE1sp1A穿梭质粒和pJM 17 腺病毒质粒，3）用重组腺病毒载体构建重组腺病毒。 在293个肾细胞中进行同源重组程序，4）繁殖 重组腺病毒在293细胞中进行大规模纯化， 5)通过在293细胞中的噬斑测定来滴定纯化的腺病毒。核心 还将负责重组腺病毒的长期储存 并将协助个体研究者进行基因转移 方案在成人心肌细胞中。

The function of the Gene Transfer Core is to provide a facility for the construction, screening, large-scale purification, and tittering 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 293 kidney cells in culture, 2) amplify and maintain purified stocks of the pdeltaE1sp1A shuttle plasmid and the pJM17 adenovirus plasmid, 3) construct recombinant adenoviruses by the homologous recombination procedure in 293 kidney cells, 4) propagate recombinant adenoviruses in 293 cells with large-scale purifications, and 5) titer purified adenoviruses by plaque assays in 293 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.