GENOME AND GENETICS OF A EUKARYOTIC ALGAL VIRUS

我们最近分离并部分鉴定了四种不同的dsdna。 在与水母共生的类小球藻中复制的病毒 和草履虫。尝试感染一些培养的小球藻菌株 四种病毒揭示了其中一种病毒PBCV-1复制的病毒 在两个小球藻菌株中。这使得镁的生产 真核病毒的数量以及非生物病毒的发展 (斑块)法检测PBCV-1。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。这个 这项提案的目标是调查结构性组织 大的病毒基因组(约280kbp)以及病毒的性质和功能 病毒基因产品。利用既定的方法，我们打算：(I) 描述dsDNA基因组，(Ii)尝试感染藻类原生质体 用分离的PBCV-1 DNA，(Iii)准备基因组的限制图， (4)监测病毒过程中病毒基因产物的合成 复制周期，以及(V)分离和鉴定温度敏感性 PBCV-1的突变体。这项研究有几个潜在的重要的长期 范围影响。关于PBCV-1的研究将：(I)提供机会 研究一种新型的病毒-宿主关系，(二)确定其作用 在共生中的作用，(Iii)展示了PBCV-1或PBCV-1的有用性 DNA作为将基因转移到其他藻类或高等植物中的载体， 以及(Iv)确定PBCV-1或PBCV-1裂解物是否含有新的来源 植物细胞壁降解酶。最后对监管和监管进行了研究。 DsDNA病毒基因组在宿主中的表达将提供新的 真核植物中基因调控的信息。

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.