TYPE 12 ADENOVIRUS TRANSFORMATION - DEFECTIVE MUTANTS

12 型腺病毒转化 - 有缺陷的突变体

• 批准号: 3170822
• 负责人: JAMES F WILLIAMS
• 金额: $ 14.34万
• 依托单位: CARNEGIE-MELLON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-08-01 至 1988-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3170822
• 关键词: Adenoviridae; defective virus; gene complementation; genetic manipulation; kidney cell; monoclonal antibody; mutant; oncogenic virus; tissue /cell culture; viral carcinogenesis; virus RNA; virus antigen; virus assembly; virus cytopathogenic effect; virus replication

The overall aim of this project is to use genetic and molecular approaches to define the functions that the various type 12 E1 proteins play in transforming mouse and rat cells in vitro, and to understand the basis for the striking differences in viral oncogenicity between this serotype and non-oncogenic type 5. Our specific aims over the next three years are: 1) To extend genetic and biochemical characterization of the host-range (hr) mutants of type 12 which we have recently isolated. 2) To isolate and characterize conditional transformation-defective mutants of this serotype. 3) To isolate and characterize site-specific mutants of type 12, particularly those which alter E1A mRNA splice junctions. 4) To isolate type 12 transformants of A549 cells. 5) To analyze the transformation process in mouse kidney cells, and determine if epithelial and fibroblastic subpopulations are transformed at different frequencies. 6) Extend our study of viral oncogenicity and cellular malignancy by testing the ability of hr mutants to induce tumors and tumor specific transplantation immunity. 7) Using type 12 hr mutants, develop a complementing system for isolation and growth of non-conditionally defective mutants.

该项目的总体目的是使用遗传和分子方法 定义各种12 E1蛋白在 在体外转化小鼠和大鼠细胞，并了解 这种血清型和 非癌性5型。我们未来三年的具体目标是：1） 扩展宿主范围（HR）的遗传和生化表征 我们最近分离的12型突变体。 2）分离和 表征有条件转化的缺陷突变体 血清型。 3）分离并表征12型的位点特异性突变体， 特别是那些改变E1A mRNA剪接连接的人。 4）分离 A549细胞的12型转化物。 5）分析转换 小鼠肾细胞中的过程，并确定上皮和成纤维细胞是否 亚群以不同的频率转换。 6）扩展我们的 通过测试能力来研究病毒致癌性和细胞恶性肿瘤 人力资源突变体诱导肿瘤和肿瘤特异性移植 免疫。 7）使用12型小时突变体，开发一个补充系统 非条件有缺陷突变体的隔离和生长。