RNAs transcribed from a 3.6-kilobase SmaI fragment of the short unique region of the herpes simplex virus type 1 genome.
RNAs transcribed from a 3.6-kilobase SmaI fragment of the short unique region of the herpes simplex virus type 1 genome.
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RNA 转录自 1 型单纯疱疹病毒基因组的短独特区域的 3.6 KB SmaI 片段。
DOI：
10.1128/jvi.48.2.460-471.1983
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发表时间：
1983
影响因子：
5.4
通讯作者：
Pizer,LI
中科院分区：
文献类型：
--
作者：
Ikura,K;Betz,JL;Sadler,JR;Pizer,LI
利用单纯疱疹病毒1型(Kos)DNA BamHI J片段的3.6kb SmaI亚克隆，对编码糖蛋白D的基因组片段(0.91~0.93map单位)转录的mRNAs进行了鉴定。RNA印迹显示有两个主要的RNA片段，分别为2.3和1.5kb。与32P末端标记的DNA片段的5‘和3’定位表明，这些RNA是一个嵌套的集合，每个RNA都有自己的启动子和3‘末端。此外，还观察到5‘末端不连续的RNA种类较少。精确的5‘定位和测序数据定位了起始点，并在适当的位置显示了TATA盒、CAT盒和AC富集区。3‘作图为两个mRNAs定位了一个共同的末端，但通过在RNA末端附近的一点剪接，2.3kb的mRNAs的尺寸减小了。体外径流转录实验证实了这两个启动子的位置，并表明未感染的细胞提取物在这两个位置都忠实地启动了。尽管DNA结构相似，体外利用启动子的效率明显相同，但2.3-kb的mRNA在感染后1h就出现在细胞质中，而1.5-kb的mRNA则推迟到感染后3h才出现。
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.