Mullaney J, Moss H W, McGeoch D J
Institute of Virology, University of Glasgow, U.K.
J Gen Virol. 1989 Feb;70 ( Pt 2):449-54. doi: 10.1099/0022-1317-70-2-449.
An insertion mutant of herpes simplex virus type 1 has been constructed which carries the lacZ gene from Escherichia coli within the coding sequence of gene UL2, which is in the long unique region of the genome. In a one-step growth curve experiment this recombinant (called in 1601) grew as well as the wild-type (wt) parent virus, indicating that the UL2 gene is dispensable for growth in tissue culture. Analysis of in 1601 DNA with restriction endonucleases showed no detectable changes from the wt apart from the insertion. Extracts of cells infected with in 1601 possessed levels of viral DNA polymerase and alkaline exonuclease activities similar to those infected with the wt, but unlike the wt had negligible uracil-DNA glycosylase activity, suggesting strongly that the product of the UL2 gene is the uracil-DNA glycosylase. The sequence of the uracil-DNA glycosylase gene of E. coli was recently published, and the encoded amino acid sequence of this shows clear similarity to that of UL2, confirming our results.
已构建出1型单纯疱疹病毒的一个插入突变体,它在基因组的长独特区域内的基因UL2编码序列中携带来自大肠杆菌的lacZ基因。在一步生长曲线实验中,这种重组体(称为1601)的生长情况与野生型(wt)亲本病毒一样好,这表明UL2基因对于在组织培养中的生长是可有可无的。用限制性内切酶分析1601的DNA表明,除了插入之外,与野生型没有可检测到的变化。感染1601的细胞提取物中病毒DNA聚合酶和碱性外切核酸酶活性水平与感染野生型的细胞相似,但与野生型不同的是,其尿嘧啶-DNA糖基化酶活性可忽略不计,这强烈表明UL2基因的产物就是尿嘧啶-DNA糖基化酶。大肠杆菌尿嘧啶-DNA糖基化酶基因的序列最近已发表,并显示其编码的氨基酸序列与UL2的氨基酸序列有明显相似性,证实了我们的结果。