Bogner E, Reschke M, Reis B, Mockenhaupt T, Radsak K
Institut für Virologie der Philipps-Universität, Marburg, Germany.
Virology. 1993 Sep;196(1):290-3. doi: 10.1006/viro.1993.1477.
Experiments were undertaken to identify the product of open reading frame (orf) UL56 of human cytomegalovirus (HCMV), the putative homolog of infected cell protein (ICP) 18.5 of herpes simplex virus type 1 (HSV-1) that is thought to be involved in viral nucleocapsid maturation. Northern blotting using a unique fragment of orf UL56 revealed a specific transcript of about 3.0 kb in HCMV-infected fibroblasts early and late postinfection (p.i.). Two overlapping fragments of UL56 were subcloned for procaryotic expression and the recombinant proteins were used for affinity purification of an antibody fraction from human convalescent serum (pab UL56). The affinity purified antibody recognized a polypeptide with an apparent molecular weight of about 130 kDa (p 130) in immunoblots of virions and of infected cell extracts late p.i. Upon treatment of isolated virions with nonionic detergent the p 130 polypeptide separated with the nucleocapsid/tegument fraction. The amino acid sequence of p 130 deduced from the nucleotide sequence exhibited a high homology to ICP 18.5 of HSV-1.
开展了实验以鉴定人巨细胞病毒(HCMV)开放阅读框(orf)UL56的产物,它被认为是单纯疱疹病毒1型(HSV-1)感染细胞蛋白(ICP)18.5的同源物,且被认为参与病毒核衣壳成熟过程。使用orf UL56的一个独特片段进行Northern印迹分析,发现在HCMV感染的成纤维细胞感染后早期和晚期(p.i.)有一条约3.0 kb的特异性转录本。将UL56的两个重叠片段亚克隆用于原核表达,重组蛋白用于从人恢复期血清中亲和纯化抗体组分(pab UL56)。亲和纯化的抗体在病毒粒子和感染后晚期的感染细胞提取物的免疫印迹中识别出一种表观分子量约为130 kDa的多肽(p 130)。用非离子去污剂处理分离的病毒粒子后,p 130多肽与核衣壳/包膜组分分离。从核苷酸序列推导的p 130氨基酸序列与HSV-1的ICP 18.5具有高度同源性。