MOLECULAR GENETICS OF VARICELLA-ZOSTER VIRUS

水痘-带状疱疹病毒（VZV）是水痘的病原体， 带状疱疹。 我们已经开发了一个系统，以产生重组VZV 通过用四个重叠的VZV粘粒分离组织培养细胞， DNA 突变已经被工程化到导致VZV的cosplay中， 不能表达病毒胸苷酸合成酶，糖蛋白V， 或核糖核苷酸还原酶基因。 无法表达的病毒 前两个基因的生长速度与亲本病毒相似， 不能表达核糖核苷酸还原酶基因的病毒 以比野生型病毒更慢的速度传播。 将使用选定的VZV突变体 给豚鼠注射疫苗，以确定病毒是否失去了 建立中枢神经系统潜伏期的能力。 外源基因已被插入VZV基因组中。 急诊大肠杆菌 半乳糖苷酶基因，已被插入VZV基因组， 所得病毒产生用Xgal染成蓝色的噬斑。 这种病毒 已经被接种到豚鼠身上，以确定细胞的类型， 能够支持病毒复制和潜伏期的建立。 的 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和产生的病毒 在感染细胞表面表达高水平的HSV gD。 将用表达HSV gD的VSV接种动物，以确定是否 它们被保护免受HSV感染。 已经开发了分泌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并用活VZV攻击，以确定它们是否 都不会被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.