Ikura K, Betz J L, Sadler J R, Pizer L I
J Virol. 1983 Nov;48(2):460-71. doi: 10.1128/JVI.48.2.460-471.1983.
A 3.6-kilobase (kb) SmaI subclone of the BamHI J fragment of herpes simplex virus type 1 (KOS) DNA was utilized to characterize the mRNAs transcribed from the genome segment (0.91 to 0.93 map units) that encodes glycoprotein D mRNA. RNA blotting demonstrated two major RNA species of 2.3 and 1.5 kb. 5' and 3' mapping with 32P-end-labeled DNA fragments indicated that these RNAs are a nested set, each having its own promoter and 3' terminus. Less abundant RNA species with discrete 5' ends were also observed. Precise 5' mapping and sequence data located the initiation sites and demonstrated TATA boxes, CAT boxes, and AC-rich regions in the appropriate positions. 3' mapping located a common end for both mRNAs, but the 2.3-kb mRNA was reduced in size by splicing at a point near the RNA terminus. In vitro runoff transcription experiments confirmed the location of the two promoters and showed that an uninfected cell extract initiated faithfully at both sites. Despite the similarities in DNA structure and the apparent equal efficiency of promoter utilization in vitro, the 2.3-kb mRNA appeared in the cytoplasm early (1 h) after infection, whereas the 1.5-kb mRNA was delayed until 3 h after infection.
利用单纯疱疹病毒1型（KOS）DNA的BamHI J片段的一个3.6千碱基（kb）的SmaI亚克隆来表征从编码糖蛋白D mRNA的基因组片段（0.91至0.93图谱单位）转录的mRNA。RNA印迹显示有两种主要的RNA，大小分别为2.3 kb和1.5 kb。用32P末端标记的DNA片段进行5'和3'定位表明，这些RNA是一组嵌套的RNA，每个都有自己的启动子和3'末端。还观察到具有离散5'末端的丰度较低的RNA种类。精确的5'定位和序列数据确定了起始位点，并在适当位置显示了TATA框、CAT框和富含AC的区域。3'定位确定了两种mRNA的共同末端，但2.3 kb的mRNA在RNA末端附近的一个点通过剪接而减小了大小。体外径流转录实验证实了两个启动子的位置，并表明未感染细胞提取物在两个位点都能准确起始。尽管DNA结构相似且体外启动子利用效率明显相同，但2.3 kb的mRNA在感染后早期（1小时）出现在细胞质中，而1.5 kb的mRNA则延迟到感染后3小时出现。
