Stevens J G, Wagner E K, Devi-Rao G B, Cook M L, Feldman L T
Science. 1987 Feb 27;235(4792):1056-9. doi: 10.1126/science.2434993.
In initial attempts to define the molecular events responsible for the latent state of herpes simplex virus, in situ hybridization was utilized to search for virally encoded RNA transcripts in latently infected sensory neurons. The use of cloned probes representing the entire viral genome indicated that transcripts encoded within terminal repeats were present. When the alpha genes encoding ICP-0, ICP-4, and ICP-27 and the gamma 1 gene encoding VP-5 were employed, only RNA transcripts hybridizing to the ICP-0 probe were detected. In latently infected cells, the ICP-0--related transcripts were localized principally in the nucleus; this was not the case in acutely (productively) infected neurons or in neurons probed for RNA transcripts coding for actin. In Northern blotting experiments, an RNA of 2.6 kilobases was detected with the ICP-0 probe. When single-stranded DNAs from the ICP-0 region were used as probes, RNA from the strand complementary to that encoding ICP-0 messenger RNA (mRNA) was the major species detected. This RNA species may play a significant role in maintaining the latent infection.
在最初尝试确定导致单纯疱疹病毒潜伏状态的分子事件时,采用原位杂交技术在潜伏感染的感觉神经元中寻找病毒编码的RNA转录本。使用代表整个病毒基因组的克隆探针表明,末端重复序列内编码的转录本存在。当使用编码ICP-0、ICP-4和ICP-27的α基因以及编码VP-5的γ1基因时,仅检测到与ICP-0探针杂交的RNA转录本。在潜伏感染的细胞中,与ICP-0相关的转录本主要定位于细胞核;在急性( productive)感染的神经元或检测肌动蛋白编码RNA转录本的神经元中则并非如此。在Northern印迹实验中,用ICP-0探针检测到一条2.6千碱基的RNA。当使用来自ICP-0区域的单链DNA作为探针时,与编码ICP-0信使RNA(mRNA)的链互补的链上的RNA是检测到的主要种类。这种RNA种类可能在维持潜伏感染中起重要作用。