戊型肝炎病毒亚基因组RNA的研究

STUDIES ON THE SUBGENOMIC RNAS OF HEPATITIS E VIRUS

本文利用同位素代谢标记在HEV感染 8 5～ 1 0 5,6 5～ 7 5h分别检测到 1及 2个亚基因组RNA ,而感染 2 1h后及在成熟的病毒颗粒内未能检测到亚基因组RNA。通过杂交实验 ,发现HEV的亚基因组RNA具有典型的共 3′端的半套式结构 ,且基因组RNA与亚基因组RNA的 5′端不存在共同的引导序列。通过紫外转录图谱发现HEV的亚基因组RNA是通过独立转录的方式产生的。利用引物延伸反应发现两种亚基因组RNA的转录起始位点分别位于RNA聚合酶区及非结构区、结构区的基因间序列。

Through labeling HEV RNAs with [ 32 P] phosphate, two subgenomic RNAs at 6 5 to 7 5h post infection, one subgenomic RNA at 8 5 to 10 5h post infection and none subgenomic RNAs were detected at 21 post infection or in purified virus. Hybridization results illustrated that the HEV subgenomic RNAs form a 3′ nested set which is situated at the 3′ end of genome, but their 5′ ends do not share a common leader sequence. UV transcription results showed that the HEV subgenomic RNAs were transcribed independently from its promote. Primer extention analysis sugested that the 5′ end of large subgenomic RNA was mapped to the initiative region within RNA polymerase region, and the small sugenomic RNA was mapped to the intergenic region between ORF1 and ORF2.