STRUCTURAL ANALYSIS OF THE GENOME OF RESPIRATORY SYNCYTIAL VIRUS

呼吸道合胞病毒基因组的结构分析

• 批准号: 3746526
• 负责人: P L COLLINS
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3746526
• 关键词: chloramphenicol acetyltransferase; complementary DNA; genetic regulatory element; genome; microorganism culture; molecular cloning; respiratory syncytial virus; viral rescue; virus RNA; virus genetics; virus replication; viruslike particle

Previously, cDNAs were constructed to encode various truncated versions of the genome RNA of respiratory syncytial virus (RSV) in which the genome termini and putative transcription signals were retained but the viral genes were deleted and replaced with the reporter gene for bacterial chloramphenicol acetyl transferase (CAT). When introduced into RSV-infected cells, the prototype RSV-CAT minigenome appeared to be replicated, transcribed and packaged into virus-like particles. This represents the first type of system in which genome-like RNAs of a nonsegmented negative strand RNA virus were introduced into the viral replication cycle and rendered biologically active. It is an important step in developing the technology for rescuing complete replication- competent virus from cDNA. This capability would allow the engineering of defined viruses as attenuated vaccines and as reagents for molecular biological and viral pathogenesis studies. In its present form, the RSV- CAT system was used to identify and characterize, for the first time, the cis-acting replication and transcription signals in the RSV genome. It also should be possible to adapt the RSV-CAT system for the analysis of protein function (accompanying report).

以前，构建了cDNA以编码各种截短版本 呼吸综合病毒（RSV）的基因组RNA，其中 保留了基因组末端和推定的转录信号 将病毒基因删除并用报告基因代替 细菌氯霉素乙酰转移酶（CAT）。当引入 RSV感染的细胞，原型RSV-CAT微型素组似乎是 复制，转录并包装成病毒样颗粒。这 代表了第一种类型的系统，其中类似于基因组的RNA 将未分段的阴性链RNA病毒引入了病毒 复制周期并具有生物学活性。这是一个重要的 开发用于营救完整复制的技术的一步 来自cDNA的胜任病毒。这种能力将允许工程 定义的病毒作为减毒疫苗和分子试剂的试剂 生物学和病毒发病机理研究。以目前的形式，RSV- 猫系统被用来识别和表征 RSV基因组中的顺式作用复制和转录信号。它 也应该可以调整RSV-cat系统以分析 蛋白质功能（随附的报告）。