GENOME AND GENETICS OF A EUKARYOTIC ALGAL VIRUS

真核藻类病毒的基因组和遗传学

• 批准号: 3281269
• 负责人: JAMES L VAN ETTEN
• 金额: $ 10.12万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-09-28 至 1987-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3281269
• 关键词: Chlorophyta; DNA virus; eukaryote; gene expression; genetic mapping; genetic regulation; genome; plant genetics; plant virus; plaque assay; temperature sensitive mutant; virus DNA; virus RNA; virus genetics; virus replication

We have recently isolated and partially characterized four distinct dsDNA viruses which replicate in Chlorella-like green algae symbiotic with Hydra and Paramecium. Attempts to infect a number of cultured Chlorella strains with the four viruses revealea that one of the viruses, PBCV-1, replicated in two of the Chlorella strains. This has allowed the production of mg quantities of a eukaryotic virus as well as the development of abiological (plaque) assay for PBCV-1. The PBCV-1 Chlorella system is the first example of a virus infecting any eukaryotic plant which can utilize procedures directly adapted from those used to study bacteriophage. The objectives of this proposal are to investigate the structural organization of the large viral genome (ca. 280 kbp) and the nature and function of the viral gene products. Using established methods we intend to (i) characterize the dsDNA genome, (ii) attempt to infect algal protoplasts with isolated PBCV-1 DNA, (iii) prepare a restriction map of the genome, (iv) monitor the synthesis of viral gene products during the viral replication cycle, and (v) isolate and characterize temperature sensitive mutants of PBCV-1. The research has several potentially important long range implications. Studies on PBCV-1 will: (i) provide an opportunity to study a new type of virus-host relationship, (ii) determine the role it plays in symbiosis, (iii) demonstrate the usefulness of PBCV-1 or PBCV-1 DNA as a vector for transferring genes into other algae or higher plants, and (iv) determine if PBCV-1 or PBCV-1 lysates contain a new source of plant cell wall degrading enzymes. Finally studies on the regulation and expression of the dsDNA viral genome in the host will provide new information on gene regulation in eukaryotic plants.

我们最近分离并部分表征了四种不同的dsDNA 在与水螅共生的类似小球藻的绿色藻类中复制的病毒 和草履虫 尝试感染许多培养的小球藻菌株 这四种病毒显示其中一种病毒PBCV-1复制了 在两个小球藻菌株中。 这使得mg的生产 大量的真核病毒以及非生物学的发展 针对PBCV-1的噬菌斑（空斑）测定。 PBCV-1小球藻系统是第一个 感染任何可以利用的真核植物的病毒的例子 程序直接改编自用于研究噬菌体的程序。 的 本提案的目的是调查结构组织 大的病毒基因组（ca. 280 kbp）的性质和功能 病毒基因产物 使用已建立的方法，我们打算（i） 表征dsDNA基因组，（ii）尝试感染藻类原生质体 用分离的PBCV-1 DNA，（iii）制备基因组的限制性图谱， (iv)在病毒感染期间监测病毒基因产物的合成， 复制周期，和（v）分离和表征温度敏感 PBCV-1的突变体。 这项研究有几个潜在的重要长期 范围影响。 对PBCV-1的研究将：㈠提供机会， 研究一种新型的病毒-宿主关系，（ii）确定它的作用， （iii）证明PBCV-1或PBCV-1的有用性 DNA作为将基因转移到其它藻类或高等植物中的载体， 和（iv）确定PBCV-1或PBCV-1裂解物是否含有新的 植物细胞壁降解酶 最后，研究了管制和 dsDNA病毒基因组在宿主中的表达将提供新的 真核植物基因调控的信息。