Carmichael E P, Weller S K
Department of Microbiology, University of Connecticut Health Center, Farmington 06032.
J Virol. 1989 Feb;63(2):591-9. doi: 10.1128/JVI.63.2.591-599.1989.
We investigated the role of the herpes simplex virus type 1 UL8 gene product in viral DNA replication. First, we unambiguously fine mapped the mutation in tsS38 (complementation group 1-26) to an open reading frame, designated UL8, predicted to encode an 80-kilodalton protein. Previous studies indicated that tsS38 was capable of synthesizing low to moderate levels of viral DNA at the nonpermissive temperature (C. T. Chu, D. S. Parris, R. A. F. Dixon, F. E. Farber, and P. A. Schaffer, Virology 98:168-181, 1979); thus, it was not clear whether the UL8 gene product is essential for viral DNA synthesis. Therefore, a deletion-insertion mutation was constructed in the UL8 gene by removing most of its coding sequences and replacing them with the Escherichia coli lacZ gene under control of the viral ICP6 regulatory signals. The resulting recombinant, hr80, was propagated in helper cells (S22) which express the wild-type version of the UL8 gene, but was incapable of forming plaques in Vero cells. Furthermore, hr80 was totally defective in the synthesis of viral DNA and late proteins under nonpermissive growth conditions. These results demonstrated that the UL8 gene product is essential for viral DNA synthesis.
我们研究了单纯疱疹病毒1型UL8基因产物在病毒DNA复制中的作用。首先，我们明确地将tsS38（互补组1 - 26）中的突变精细定位到一个开放阅读框，命名为UL8，该开放阅读框预计编码一种80千道尔顿的蛋白质。先前的研究表明，tsS38在非允许温度下能够合成低至中等水平的病毒DNA（C.T.朱、D.S.帕里斯、R.A.F.迪克森、F.E.法伯和P.A.沙弗，《病毒学》98:168 - 181，1979）；因此，尚不清楚UL8基因产物对于病毒DNA合成是否必不可少。所以，通过去除UL8基因的大部分编码序列并用病毒ICP6调控信号控制下的大肠杆菌lacZ基因进行替换，构建了一个缺失 - 插入突变体。所得的重组体hr80在表达野生型UL8基因的辅助细胞（S22）中增殖，但在Vero细胞中无法形成噬斑。此外，在非允许生长条件下，hr80在病毒DNA和晚期蛋白的合成方面完全存在缺陷。这些结果表明，UL8基因产物对于病毒DNA合成是必不可少的。
