Desloges N, Boucher H, Simard C
Centre de Microbiologie et Biotechnologie, INRS-Institut Armand-Frappier, Québec, Canada.
Arch Virol. 2001;146(9):1693-704. doi: 10.1007/s007050170057.
We investigated whether the bovine herpesvirus 1.1 (BHV1) ORF, a homologue of the herpes simplex virus 1 (HSV1) UL25 gene, represented a functional gene. The BHV1 UL25 ORF, which is located at positions 60602<--62398 of the viral genome, generated a 4.5 kb transcript accumulating at low abundance as soon as 3 hours p.i. after which the levels increased up to 12h p.i. and remained constant up to 24 hours p.i. In addition, UL25 transcription initiated at 303 bases upstream from the translation initiation codon, corresponding to 26 and 354 b downstream from putative TATA and CAAT boxes, respectively, thus providing evidence that these elements function as the UL25 promoter. Western blotting of BHV1-infected cell lysates, using a BHV1-UL25 specific antiserum generated against a T7-Tag/UL25 fusion recombinant protein expressed in E. coli, detected a 63 kDa protein of the expected size (63,083 Da) whose expression profile followed that of its transcript. The synthesis of the 63 kDa protein was abrogated by a DNA synthesis inhibitor, unambiguously demonstrating that the viral specific protein expressed from the BHV1 UL25 ORF belongs to the gamma2 class.
我们研究了牛疱疹病毒1.1（BHV1）的开放阅读框（ORF），它是单纯疱疹病毒1（HSV1）UL25基因的同源物，是否代表一个功能基因。BHV1 UL25 ORF位于病毒基因组的60602<--62398位，产生一个4.5 kb的转录本，感染后3小时就开始以低丰度积累，之后其水平在感染后12小时上升，并在感染后24小时保持恒定。此外，UL25转录起始于翻译起始密码子上游303个碱基处，分别对应于假定的TATA盒和CAAT盒下游26和354个碱基，从而证明这些元件作为UL25启动子发挥作用。使用针对在大肠杆菌中表达的T7标签/UL25融合重组蛋白产生的BHV1-UL25特异性抗血清，对BHV1感染的细胞裂解物进行蛋白质免疫印迹分析，检测到一种预期大小（63,083 Da）的63 kDa蛋白，其表达谱与其转录本的表达谱一致。一种DNA合成抑制剂消除了该63 kDa蛋白的合成，明确表明从BHV1 UL25 ORF表达的病毒特异性蛋白属于γ2类。
