Hamel Francine, Boucher Hélène, Simard Claire
INRS-Institut Armand-Frappier, 531 Boulevard des Prairies, Laval-des-Rapides, Quebec, Canada H7V 1B7.
Virus Res. 2002 Mar 20;84(1-2):125-34. doi: 10.1016/s0168-1702(02)00002-3.
We characterized the expression kinetics of the transcript and protein generated from the bovine herpesvirus 1 (BHV1) homologue of the herpes simplex virus 1 (HSV1) UL51 gene. The BHV1 UL51 ORF, located at positions 7236-->7967 of the viral genome, generated a major 1.05 kb transcript accumulating at very low abundance as soon as 3 h post-infection (p.i.), after which its levels increased to reach a plateau from 6 to 12 h p.i., and then slowly decreased up to 24 h p.i. As determined by S1 nuclease protection assays, UL51 transcription initiated at two distinct sites located at 191 and 196 bases upstream from the initiation codon, corresponding to positions 7045 and 7040 of the viral genome, respectively. Western blotting of BHV1-infected protein cell lysates, using a BHV1-specific antiserum generated against a recombinant protein expressed in Escherichia coli, detected a 28 kDa protein of the expected size (24985 Da) whose expression kinetics followed that of its transcript. As evidenced by in situ immunofluorescence assays, the protein mainly localized to the cytoplasm and the perinuclear region of infected cells. In contrast to HSV1 UL51 which is classified as a gamma2 gene, BHV1 UL51 belongs to viral genes of the gamma1 class as expression of its transcript is partially dependent on viral DNA synthesis.
我们对单纯疱疹病毒1型（HSV1）UL51基因的牛疱疹病毒1型（BHV1）同源物所产生的转录本和蛋白质的表达动力学进行了表征。BHV1 UL51开放阅读框位于病毒基因组的7236→7967位，产生了一个主要的1.05 kb转录本，感染后3小时（p.i.）即开始以非常低的丰度积累，此后其水平在感染后6至12小时上升至平台期，然后在感染后24小时缓慢下降。通过S1核酸酶保护试验确定，UL51转录起始于起始密码子上游191和196个碱基处的两个不同位点，分别对应于病毒基因组的7045和7040位。使用针对在大肠杆菌中表达的重组蛋白产生的BHV1特异性抗血清对BHV1感染的蛋白质细胞裂解物进行蛋白质印迹分析，检测到一种预期大小（24985 Da）的28 kDa蛋白质，其表达动力学与其转录本的表达动力学一致。原位免疫荧光试验表明，该蛋白质主要定位于受感染细胞的细胞质和核周区域。与被归类为γ2基因的HSV1 UL51不同，BHV1 UL51属于γ1类病毒基因，因为其转录本的表达部分依赖于病毒DNA合成。
