Chang Y E, Roizman B
Marjorie B. Kovler Viral Oncology Laboratories, University of Chicago, Illinois 60637.
J Virol. 1993 Nov;67(11):6348-56. doi: 10.1128/JVI.67.11.6348-6356.1993.
The nucleotide sequence of the UL31 open reading frame is predicted to encode a basic protein with a hydrophilic amino terminus and a nuclear localization signal. To identify its gene product, we constructed a viral genome in which the thymidine kinase gene was inserted between the UL31 and UL32 open reading frames. The thymidine kinase gene was then deleted, and in the process, the 5' terminus of the UL31 open reading frame was replaced with a 64-bp sequence in frame with the complete, authentic sequence of the UL31 open reading frame. The inserted sequence encoded a hydrophilic epitope derived from glycoprotein B of human cytomegalovirus and for which a monoclonal antibody is available. We report that in infected cells, the tagged protein localized in and was dispersed throughout the nucleus. Nuclear fractionation studies revealed that the UL31 protein partitions with the nuclear matrix. The protein is phosphorylated in infected cells maintained in medium containing 32Pi.
预测UL31开放阅读框的核苷酸序列编码一种具有亲水性氨基末端和核定位信号的碱性蛋白。为了鉴定其基因产物，我们构建了一个病毒基因组，其中胸苷激酶基因插入在UL31和UL32开放阅读框之间。然后删除胸苷激酶基因，在此过程中，UL31开放阅读框的5'末端被一个64碱基对的序列取代，该序列与UL31开放阅读框的完整、真实序列读码框一致。插入序列编码一个源自人巨细胞病毒糖蛋白B的亲水性表位，并且有针对该表位的单克隆抗体。我们报道，在受感染细胞中，标记蛋白定位于细胞核并分散于整个细胞核。细胞核分级分离研究表明，UL31蛋白与核基质共分离。在含有32Pi的培养基中培养的受感染细胞中，该蛋白发生磷酸化。
