Jiang Y M, Yamada H, Goshima F, Daikoku T, Oshima S, Wada K, Nishiyama Y
Laboratory of Virology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine, Japan.
J Gen Virol. 1998 Nov;79 ( Pt 11):2777-84. doi: 10.1099/0022-1317-79-11-2777.
The herpes simplex virus type 2 (HSV-2) US2 gene product was identified by using a rabbit polyclonal antiserum raised against a recombinant 6 x His-US2 fusion protein expressed in Escherichia coli. The antiserum reacted specifically with a 39 kDa protein in HSV-2 strain 186-infected cell lysates. The protein was not detectable in the presence of the virus DNA synthesis inhibitor phosphonoacetic acid. Indirect immunofluorescence studies localized the US2 protein in the cytoplasm and as discrete granules at late times post-infection within and at the periphery of the nucleus, and nuclear fractionation studies showed that the protein was partially associated with the nuclear matrix of infected cells. The protein was easily detected in purified virions. Also, a US2 insertion mutant was constructed which contained an ICP6-lacZ insertion in the US2 gene. This mutant was as virulent as wild-type virus in mice when inoculated by the footpad route. The importance of the US2 protein of HSV-2 in the virus life-cycle may be apparent only in the natural human host.
通过使用针对在大肠杆菌中表达的重组6×His-US2融合蛋白产生的兔多克隆抗血清，鉴定出了2型单纯疱疹病毒（HSV-2）的US2基因产物。该抗血清与HSV-2 186株感染的细胞裂解物中的一种39 kDa蛋白发生特异性反应。在存在病毒DNA合成抑制剂膦甲酸的情况下，无法检测到该蛋白。间接免疫荧光研究表明，US2蛋白定位于细胞质中，在感染后期位于细胞核内和核周边呈离散颗粒状，细胞核分级分离研究表明该蛋白部分与感染细胞的核基质相关。在纯化的病毒粒子中很容易检测到该蛋白。此外，构建了一个US2插入突变体，该突变体在US2基因中含有一个ICP6-lacZ插入。当通过足垫途径接种时，该突变体在小鼠中的毒力与野生型病毒相同。HSV-2的US2蛋白在病毒生命周期中的重要性可能仅在天然人类宿主中才明显。
