Maray T, Malkinson M, Becker Y
Department of Molecular Virology, Faculty of Medicine, Hebrew University, Jerusalem, Israel.
Virus Genes. 1988 Oct;2(1):49-68. doi: 10.1007/BF00569736.
RNA was isolated from two strains of Marek's disease virus (MDV-Z and MDV-B). The virus was grown in duck embryo fibroblasts (DEF) for 96 hr, 72 hr in the presence of phosphonoacetic acid (PAA) and 24 hr in the presence of cycloheximide added at the time of infection. With the use of DNA probes representing about 80% of the MDV genome, an extensive Northern blot analysis of the RNA was carried out. A similar analysis was done with RNA extracted from the MDV-transformed cell line MSB-1. This study revealed 42, 25 and 29 discrete viral RNA transcripts in MDV-Z and MDV-B-infected DEF and in the MSB-1 cell line, respectively, ranging in size from 0.8 to 13 kb. In MDV-Z-infected DEF, there were twelve late RNA species, two early and eight immediate-early viral transcripts. In MDV-B-infected DEF there were eleven late RNA species, two early and seven immediate-early viral transcripts. The RNA species were homologous for all the probes used except the BamHI-G DNA fragment where no RNA transcripts were detected in the MSB-1 cell line. The RNA transcripts were used to produce a preliminary viral RNA map. Comparison of the location and sizes of the viral RNA transcripts in MDV-infected and MDV-transformed cells revealed several differences.
从两株马立克氏病病毒(MDV-Z和MDV-B)中分离出RNA。病毒在鸭胚成纤维细胞(DEF)中培养96小时,在膦甲酸(PAA)存在的情况下培养72小时,并在感染时添加环己酰亚胺的情况下培养24小时。使用代表约80%MDV基因组的DNA探针,对RNA进行了广泛的Northern印迹分析。对从MDV转化的细胞系MSB-1中提取的RNA进行了类似的分析。这项研究分别在MDV-Z和MDV-B感染的DEF以及MSB-1细胞系中揭示了42、25和29个离散的病毒RNA转录本,大小从0.8到13 kb不等。在MDV-Z感染的DEF中,有12种晚期RNA种类、2种早期和8种极早期病毒转录本。在MDV-B感染的DEF中,有11种晚期RNA种类、2种早期和7种极早期病毒转录本。除了BamHI-G DNA片段外,所有使用的探针检测到的RNA种类都是同源的,在MSB-1细胞系中未检测到该片段的RNA转录本。这些RNA转录本用于制作初步的病毒RNA图谱。比较MDV感染细胞和MDV转化细胞中病毒RNA转录本的位置和大小,发现了一些差异。