MOLECULAR BIOLOGY OF VARICELLA-ZOSTER VIRUS INFECTIONS

水痘带状疱疹病毒感染的分子生物学

• 批准号: 3822073
• 负责人: S E STRAUS
• 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/3822073
• 关键词: acyclovir; antiviral agents; cell transformation; chickenpox; communicable diseases; gene expression; genetic library; genetic manipulation; genetic mapping; genetic strain; genome; human tissue; latent virus infection; mutant; nucleic acid sequence; shingles; spinal trigeminal nucleus; varicella zoster virus; virus DNA; virus RNA; virus classification; virus genetics; virus infection mechanism

The major basic research focus of this laboratory involves this project, with the following goals: 1) to identify, map and characterize varicella-zoster virus genes and proteins active in latent or productive infections. 2) to define the temporal sequence of gene expression. 3) to determine the interaction of antiviral drugs with viral gene products though a molecular analysis of drug-resistant mutants. To accomplish these ends we have constructed a variety of recombinant libraries of the complete VZV genome. During the past year we have concluded studies of the finer mapping and directionality of viral transcripts. We mapped the viral IE 175 immediate early gene. We have transfected that gene into monkey cells and developed cell lines which stably express it. In the cell lines the expression of the gene appears to be autoregulated. Using assays by which we could measure the activity of viral gene promotors we detected a number of VZV genes that are transactivating and transactivatible. During the past year we have also completed sequence analysis of the thymidine kinase locus of ten VZV strains, including six that are resistant to acyclovir by virtue of mutations in that gene. We identified the nature and location of base mutations which render this gene product nonfunctional. Included in the analyses is the first acyclovir-resistant VZV strain recovered from a human. During the past year we have successfully established a highly sensitive and specific in situ hybridization system using 35S labelled synthetic RNA probes. Such probes VZV RNA expression in the trigeminal ganglia of six of 16 subjects. Continuing efforts will be directed at better mapping and characterizing the RNA expressed during VZV latency.

该实验室的主要基础研究重点涉及这一点 项目，目标如下： 1)水痘-带状疱疹病毒基因的鉴定、定位和特征 以及在潜伏性或生产性感染中具有活性的蛋白质。 2)来定义基因表达的时间顺序。 3)确定抗病毒药物与病毒基因的相互作用 通过对耐药突变体的分子分析， 为了实现这些目标，我们构建了各种 完整VZV基因组的重组文库。 期间 去年，我们完成了更精细的绘图研究， 病毒转录物的方向性。 我们绘制了病毒IE 175 即刻早期基因 我们已经把这个基因 猴细胞和稳定表达它的发达细胞系。 该基因表达的细胞系似乎 自动调节 通过检测，我们可以测量 病毒基因启动子的活性，我们检测到一些VZV 反式激活和可反式激活的基因。 期间 去年，我们还完成了 胸苷激酶基因座的10个VZV株，包括6个， 通过基因突变对阿昔洛韦产生抗药性。 我们 确定了碱基突变的性质和位置， 这个基因产物是无功能的 分析中包括 第一个从人体中回收的抗阿昔洛韦VZV毒株。 在过去的一年里，我们成功地建立了一个高度 敏感特异的35 S原位杂交系统 标记的合成RNA探针。 此类探针VZV RNA表达 在16个受试者中的6个的三叉神经节中。 继续努力 将致力于更好地绘制和表征RNA 在VZV潜伏期期间表达。