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Science 1 May 1987:
Vol. 236. no. 4801, pp. 576 - 579
DOI: 10.1126/science.3033824
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Articles

Science, Vol 236, Issue 4801, 576-579
Copyright © 1987 by American Association for the Advancement of Science

articles

Rapid identification of nonessential genes of herpes simplex virus type 1 by Tn5 mutagenesis

PC Weber, M Levine, and JC Glorioso

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.


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