Weber P C, Levine M, Glorioso J C
Science. 1987 May 1;236(4801):576-9. doi: 10.1126/science.3033824.
The large genome of herpes simplex virus type of (HSV-1) encodes at least 80 polypeptides, the majority of which have no recognized function. A subgroup of these gene products appears to be nonessential for virus replication in cell culture, but contributes to the complex life cycle of the virus in the host. To identify such functions, a simple insertional mutagenesis method has been used for selective inactivation of individual HSV-1 genes. The bacterial transposon Tn5 was allowed to insert randomly into cloned restriction fragments representing the entire short unique (US) region of the HSV-1 genome. Of the 12 open reading frames that were mutagenized with Tn5, mutant derivatives of US2, US4, and US5 were recombined into the virus. These three genes proved to be nonessential for HSV-1 replication in Vero (African Green monkey kidney) cells and the US4 gene appeared to be involved in viral pathogenesis in the central nervous system of mice. This rapid mutagenesis procedure should prove useful in exploring the entire HSV-1 genome as well as the genomes of other complex animal viruses.
单纯疱疹病毒1型（HSV-1）的大基因组编码至少80种多肽，其中大多数多肽的功能尚未明确。这些基因产物中的一个亚组似乎对病毒在细胞培养中的复制并非必需，但对病毒在宿主体内的复杂生命周期有作用。为了确定此类功能，一种简单的插入诱变方法已被用于选择性灭活单个HSV-1基因。使细菌转座子Tn5随机插入代表HSV-1基因组整个短独特（US）区域的克隆限制片段中。在用Tn5诱变的12个开放阅读框中，US2、US4和US5的突变衍生物被重组到病毒中。这三个基因被证明对HSV-1在非洲绿猴肾（Vero）细胞中的复制并非必需，并且US4基因似乎参与了小鼠中枢神经系统中的病毒致病过程。这种快速诱变程序在探索整个HSV-1基因组以及其他复杂动物病毒的基因组方面应会被证明是有用的。
