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人巨细胞病毒基因组长反向重复序列中主要基因的组织与表达

Organization and expression of the major genes from the long inverted repeat of the human cytomegalovirus genome.

作者信息

Hutchinson N I, Sondermeyer R T, Tocci M J

出版信息

Virology. 1986 Nov;155(1):160-71. doi: 10.1016/0042-6822(86)90176-5.

Abstract

The long inverted repeat (TRL:IRL) of the human cytomegalovirus (CMV) genome is a major transcription unit in productively infected human fibroblasts. Cloned DNA fragments of the CMV IRL and complementary DNA (cDNA) copies of RNAs transcribed from the TRL:IRL were used as probes in RNA filter hybridization experiments to characterize and map the RNAs transcribed from this region of the virus genome. In human fibroblasts, three poly A+ RNAs of 2.7, 2.0, and 1.2 kb were detected during the early (E) and late (L) phases of virus gene expression. Analysis of cloned cDNAs and RNA mapping studies indicate that the TRL:IRL can be divided into three transcriptionally active regions. The most highly transcribed region lies between 0.805 and 0.816 map units and encodes a major abundant poly A+ RNA of 2.7 kb that is expressed at E and L times postinfection (p.i.). The second region spans map coordinates 0.792-0.797 and encodes a 1.2-kb poly A+ RNA that is relatively low in abundance at E times p.i. but achieves nearly the same abundance as the 2.7-kb transcript at L times p.i. The third region encompasses map units 0.796-0.804 and encodes a less abundant poly A+ RNA of 2.0 kb that attains maximum expression at E times. The 1.2- and 2.7-kb RNAs are transcribed in the same direction, while the 2.0-kb RNA is transcribed in the opposite direction. The 2.7-, 2.0-, and 1.2-kb RNAs, as well as 5.7- and 1.8-kb transcripts were detected at immediate early times p.i. when human fibroblasts were treated with cycloheximide, but not in cells treated with anisiomycin.

摘要

人类巨细胞病毒(CMV)基因组的长反向重复序列(TRL:IRL)是在被有效感染的人类成纤维细胞中的一个主要转录单元。CMV IRL的克隆DNA片段以及从TRL:IRL转录的RNA的互补DNA(cDNA)拷贝,被用作RNA滤膜杂交实验中的探针,以表征和定位从病毒基因组该区域转录的RNA。在人类成纤维细胞中,在病毒基因表达的早期(E)和晚期(L)阶段检测到了三种大小分别为2.7、2.0和1.2 kb的多聚腺苷酸化(poly A+)RNA。对克隆cDNA的分析和RNA定位研究表明,TRL:IRL可分为三个转录活性区域。转录最活跃的区域位于0.805至0.816图谱单位之间,编码一种2.7 kb的主要丰富多聚A+ RNA,该RNA在感染后(p.i.)的E期和L期表达。第二个区域跨越图谱坐标0.792 - 0.797,编码一种1.2 kb的多聚A+ RNA,该RNA在感染后E期丰度相对较低,但在感染后L期达到与2.7 kb转录本几乎相同的丰度。第三个区域涵盖图谱单位0.796 - 0.804,编码一种丰度较低的2.0 kb多聚A+ RNA,其在E期达到最大表达。1.2 kb和2.7 kb的RNA沿相同方向转录,而2.0 kb的RNA沿相反方向转录。当用环己酰亚胺处理人类成纤维细胞时,在感染后即刻早期检测到了2.7 kb、2.0 kb和1.2 kb的RNA,以及5.7 kb和1.8 kb的转录本,但在用茴香霉素处理的细胞中未检测到。

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