Using both selection enrichment and site-directed mutagenesis, a herpes simplex virus type 1 (HSV-1) strain 17 genome lacking all four I sites has been generated. The site at 0.45 map units which lies within the gene coding for a polypeptide of 28 000 molecular weight was removed by selection enrichment, while the site at 0.29 map units which lies within the gene coding for glycoprotein H was removed by site-directed mutagenesis. The parental virus from which these two I sites were deleted had previously had the sites at 0.07 and 0.63 map units removed through selection enrichment. The variant devoid of I sites (X4) showed normal growth characteristics; its phenotype was normal apart from the absence of the thymidine kinase protein, which is believed to be unrelated to the loss of I sites.


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