Singh J, Wagner E K
Department of Molecular Biology and Biochemistry, University of California, Irvine 92717, USA.
Virus Genes. 1995;10(2):127-36. doi: 10.1007/BF01702593.
The use of recombinant viruses has been essential in investigation of the biology of herpes simplex virus (HSV). In this communication we describe a number of viral recombination vectors that we have generated for use in promoter structure/function analysis within the context of the HSV-1 genome. We have utilized two regions of the HSV genome that contain genes nonessential for replication in cultured cells--the glycoprotein C (gC or UL44) locus in the UL of the genome and the area encompassing the promoter and 5' portion of the latency associated transcript (LAT) within the RL factual influence on promoters due to the site of insertion. Two different kinetic promoters were analyzed, those controlling expression of the gamma UL 38 and the beta dUTPase genes, in both loci. All constructs tested displayed reporter gene mRNA expression with expected kinetics, and we conclude that there are no neighboring cryptic promoter elements that could interfere with expression studies using the vectors described.
重组病毒的使用在单纯疱疹病毒(HSV)生物学研究中至关重要。在本通讯中,我们描述了一些为在HSV - 1基因组背景下进行启动子结构/功能分析而构建的病毒重组载体。我们利用了HSV基因组的两个区域,这两个区域包含在培养细胞中复制所非必需的基因——基因组UL区的糖蛋白C(gC或UL44)基因座,以及RL区内包含潜伏期相关转录物(LAT)启动子和5'部分的区域。由于插入位点的原因,对这两个位点中控制γ UL 38和β dUTPase基因表达的两种不同动力学启动子进行了分析。所有测试的构建体均显示出具有预期动力学的报告基因mRNA表达,并且我们得出结论,不存在可能干扰使用所述载体进行的表达研究的相邻隐蔽启动子元件。