CORE--GENE TRANSFER

核心——基因转移

• 批准号: 6485287
• 负责人: PAUL J MCDERMOTT
• 金额: $ 29万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6485287
• 关键词: biomedical facility; cardiac myocytes; recombinant virus; tissue /cell culture; transfection /expression vector

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.

基因转移核心的功能是为基因转移提供设施 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 为高效基因转入成人提供可靠的方法 心肌细胞并直接改变表型。具体来说， 核心将 1) 维持培养中的 293 个肾细胞，2) 扩增并维持 pdeltaE1sp1A 穿梭质粒和 pJM17 的纯化原液 腺病毒质粒，3)构建重组腺病毒 293 个肾细胞中的同源重组程序，4) 传播 在293细胞中进行大规模纯化的重组腺病毒，以及 5)通过噬斑测定在293细胞中滴度纯化的腺病毒。核心 还将负责重组腺病毒的长期储存 并将协助个别研究者进行基因转移 成人心肌细胞中的方案。