MOLECULAR GENETICS OF VARICELLA-ZOSTER VIRUS

水痘-带状疱疹病毒(VZV)是水痘和水痘的病原体。 带状疱疹。我们已经开发了一种产生重组VZV的系统 通过将四个重叠的VZV粘粒导入组织培养细胞 DNA。 突变已经被改造成宇宙体，导致VZV 不能表达病毒胸苷合成酶，糖蛋白V， 或核糖核酸还原酶基因。不能表达的病毒 前两种基因的生长速度与亲本病毒相似，而 不能表达核糖核苷酸还原酶基因的病毒会生长 比野生型病毒的速度要慢。精选的VZV突变体将使用 给豚鼠接种疫苗，以确定病毒是否已失去 在中枢神经系统建立潜伏期的能力。 外源基因已经插入到VZV基因组中。大肠杆菌-贝塔- 半乳糖苷酶基因已被插入到VZV基因组中，并且 合成的病毒会产生X-Gal染成蓝色的斑块。这种病毒 已经被接种到豚鼠体内，以确定 能够支持病毒复制和建立延迟。这个 单纯疱疹病毒(HSV)糖蛋白D(GD)基因，编码一个主要的 中和抗原，已被插入VZV和由此产生的病毒 在感染细胞表面表达高水平的HSV gD。 动物将接种表达HSV gD的VSV以确定是否 他们受到保护，不会感染单纯疱疹病毒。 已经开发出一种分泌VZV糖蛋白II(GpII)的细胞系， 一种重要的中和抗原。豚鼠将被注射 重组VZV gpII和活体VZV攻击以确定它们是否 被保护免受VZV感染。

Varicella-zoster virus (VZV) is the etiologic agent of chickenpox and herpes zoster. We have developed a system to generate recombinant VZV by transfecting tissue culture cells with four overlapping VZV cosmid DNAs. Mutations have been engineered into the cosmids that result in VZV that is unable to express the viral thymidylate synthetase, glycoprotein V, or ribonucleotide reductase genes. Viruses that are unable to express the former two genes grow at similar rates as the parental virus, while virus that is unable to express the ribonucleotide reductase gene grows at a slower rate than wild-type virus. Selected VZV mutants will used to inoculate guinea pigs to determine if the viruses have lost the ability to establish latency in the central nervous system. Foreign genes have been inserted into the VZV genome. The E. coli beta- galactosidase gene, has been inserted into the VZV genome, and the resultant virus produces plaques that stain blue with X gal. This virus has been inoculated into guinea pigs to determine the types of cells that are able to support viral replication and establishment of latency. The herpes simplex virus (HSV) glycoprotein D (gD) gene, encoding a major neutralizing antigen, has been inserted into VZV and the resulting virus expresses high levels of HSV gD on the surface of infected cells. Animals will be inoculated with VSV expressing HSV gD to determine if they are protected against infection with HSV. A cell line has been developed that secretes VZV glycoprotein II (gpII), an important neutralizing antigen. Guinea pigs will be injected with recombinant VZV gpII and challenged with live VZV to determine if they are protected from infection with VZV.