MACROMOLECULAR SYNTHESIS IN VIRUS-INFECTED CELLS

我们将尝试回答几个问题，涉及到 呼肠孤病毒基因组中编码的与呼肠孤病毒有关的功能 乘法。它们是：a.呼肠孤病毒形态发生将在 通过分析病毒的RNA和蛋白质成分的性质 逐渐成熟的未成熟的呼肠孤病毒颗粒，从 第一批由十种单链RNA分子组成的复合体 已经组装好了。它还将在体外通过反应各种不同的 呼肠孤病毒天然蛋白与单链RNA分子的结合及分离 并对由此产生的复合体进行表征。B.天然呼肠孤病毒蛋白 将被检查是否拥有五种酶活性，这些酶是 由呼肠孤病毒核心显示。这样的蛋白质将从细菌中分离出来 由表达载体转化成呼肠孤病毒基因 克隆的。C.将尝试阐明呼肠孤病毒的功能 蛋白质通过详细检查ts突变体在 不允许的温度。10个基因中每一个都有损伤的突变体 都是可用的。乘法周期将在以下方面进行分析 呼肠孤病毒、双链RNA和呼肠孤病毒AS的合成动力学 以及呼肠孤病毒形态发生途径上的各种颗粒。 D.各种呼肠孤病毒信使RNA物种不能翻译成 同等效率。上游及周边地区的重要性 作为控制翻译效率的元素的起始密码子将被 通过引入突变、缺失和插入进行研究。 是.。三种呼肠孤病毒的进化和功能关系 血清型将通过对它们的同源基因集进行测序来研究 检验它们蛋白质的同源集合以确定三级结构的相似性 结构以及它们的抗原区和功能区的性质。 F.利巴韦林的抗呼肠孤病毒活性的方式，以及各种纯品 干扰素种类将通过确定哪些反应 它们抑制呼肠孤病毒的增殖周期。希望这些结果能够 推广应用于轮状病毒和轮状病毒的感染 医学上的重要性。

Attempts will be made to answer several questions concerning the nature of the functions encoded in the reovirus genome and concerning reovirus multiplication. They are: a. Reovirus morphogenesis will be estudied in vivo by analyzing the nature of the RNA and protein components of progressively more mature immature reovirus particles, starting with the first complexes into which the ten individual species of ssRNA molecules are assembled. It will also be studied in vitro by reacting various combinations of native reovirus proteins with ssRNA molecules and isolating and characterizing the complexes that result. b. Native reovirus proteins will be examined for possession of the five enzyme activities that are exhibited by reovirus cores. Such proteins will be isolated from bacteria transformed by expression vectors into which reovirus genes have been cloned. c. Attempts will be made to elucidate the functions of reovirus proteins by examining in detail the multiplication cycles of ts mutants at nonpermissive temperatures. Mutants with lesions in each of the ten genes are available. Multiplication cycles will be analyzed in terms of the kinetics of synthesis of ss and dsRNA species and reovirus proteins, as well as of the various particles on the pathway of reovirus morphogenesis. d. The various reovirus messenger RNA species are not translated with equal efficiency. The importance of regions upstream from and around initiation codons as elements that control efficiency of translation will e investigated by introducing mutations, deletions and insertions into them. be. The evolutionary and functional relationships of the three reovirus serotypes will be studied by sequencing cognate sets of their genes and examining cognate sets of their proteins for similarity of tertiary structure and for the nature of their antigenic and functional domains. f. The mode of the anti-reovirus activity of ribavirin, and various pure interferon species will be examined by determining which reactions of the reovirus multiplication cycle they inhibit. Hopefully, these results can be extended and applied to infections by rotaviruses and orbiviruses of medical importance.