EXPRESSION AND REGULATION OF VACCINIA VIRUS LATE GENES

负责表达的分子机制， 牛痘病毒（VV）晚期基因的调控仍然是一个谜。 这个类 VV基因，约占VV遗传潜力的一半， 只有在病毒DNA合成开始后才表达。 VV后期成绩单 来自任何一个位点的基因在大小上都是高度异质的， 起始于许多不同的5 ′位点并随机终止。 迄今为止，已经绘制和测序的少数VV晚期基因， 揭示了此前监管要素的合谋缺失 在其他真核或原核系统中被认为是必需的，甚至 VV早期基因。 因此，VV晚期基因控制区和 可能识别它们的病毒蛋白质是未知的。 由于 鉴于情况的复杂性，解决这些问题的一种方法是 选择一些感兴趣的VV晚期基因，并使它们经受强烈的 分子生物学检查 为此，一些VV基因 参与VV复制的晚期阶段[α-鹅膏蕈碱 抗性，ts 17，和六个非协调表达VV的串联阵列 晚期基因]最近已被鉴定、绘制和测序。 的 实验试图利用迄今为止获得的信息作为以下方面的基础： 1)比较动力学分析这些基因是如何以及何时 表达。 2)制备足够的免疫试剂， 编码基因产物的功能鉴定及其 参与病毒的生命周期。 3)使用定向遗传学，基因 融合和标志物拯救技术，以揭示显着的调节 每个基因的特征 4)利用足迹法和凝胶阻滞法 鉴定相互作用的病毒和/或细胞蛋白的方法 具有VV启动子和终止子区域。 预计这样的 实验将提供相当深入的机制，VV 用它来实现其复杂发展的有序表达 在被感染的细胞内。 这些信息可能有助于 设计和构建未来使用的VV重组疫苗株 传染病的传播。

The molecular mechanisms that are responsible for the expression and regulation of vaccinia virus (VV) late genes remain an enigma. This class of VV genes, which represents about half of the VV genetic potential, is expressed only after viral DNA synthesis has begun. VV late transcripts from any single locus are highly heterogeneous in size, apparently initiating from a number of distinct 5' sites and terminating randomly. The few VV late genes that have been mapped and sequenced to date, have revealed the conspicious absence of regulatory elements previously recognized as essential in other eukaryotic or prokaryotic systems, or even VV early genes. Thus, the identity of the VV late gene control regions and the viral proteins which presumably recognize them are unknown. Due to the complexity of the situation, one approach to unraveling these questions is to select a few VV late genes of interest, and to subject them to intense molecular biological scrutiny. Towards this end, a number of VV genes which participate in the late phase of VV replication [Alpha-amanitin resistance, ts17, and a tandem array of six non-coordinately expressed VV late genes] have recently been identified, mapped, and sequenced. The experiments seek to use the information obtained thus far as a basis for: 1) A comparative kinetic analysis of how and when these genes are expressed. 2) Preparation of immunological reagents sufficient to enable functional identification of the encoded gene products and how they participate in the viral life cycle. 3) Using directed-genetics, gene fusion, and marker rescue techniques to reveal the salient regulatory features of each gene. 4) Using footprinting and gel retardation methodologies to identify the viral and/or cellular proteins which interact with VV promoter and terminator regions. It is anticipated that such experiments will provide considerable insight into the mechanisms which VV employes to achieve the ordered expression of its complex developmental program within the infected cell. This information may facilitate the design and construction of future VV recombinant vaccine strains to be used for the prophylaxsis of infectious diseases.