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小鼠γ-疱疹病毒68型裂解感染的转录组概况

Transcriptome profile of murine gammaherpesvirus-68 lytic infection.

作者信息

Ebrahimi Bahram, Dutia Bernadette M, Roberts Kim L, Garcia-Ramirez Jose J, Dickinson Paul, Stewart James P, Ghazal Peter, Roy Douglas J, Nash Anthony A

机构信息

Laboratory for Clinical and Molecular Virology, The University of Edinburgh, Summerhall Square, Edinburgh EH9 1QH, UK.

Scottish Centre for Genomic Technology and Informatics, The University of Edinburgh, Summerhall Square, Edinburgh EH9 1QH, UK.

出版信息

J Gen Virol. 2003 Jan;84(Pt 1):99-109. doi: 10.1099/vir.0.18639-0.

Abstract

The murine gammaherpesvirus-68 genome encodes 73 protein-coding open reading frames with extensive similarities to human gamma(2) herpesviruses, as well as unique genes and cellular homologues. We performed transcriptome analysis of stage-specific viral RNA during permissive infection using an oligonucleotide-based microarray. Using this approach, M4, K3, ORF38, ORF50, ORF57 and ORF73 were designated as immediate-early genes based on cycloheximide treatment. The microarray analysis also identified 10 transcripts with early expression kinetics, 32 transcripts with early-late expression kinetics and 29 transcripts with late expression kinetics. The latter group consisted mainly of structural proteins, and showed high expression levels relative to other viral transcripts. Moreover, we detected all eight tRNA-like transcripts in the presence of cycloheximide and phosphonoacetic acid. Lytic infection with MHV-68 also resulted in a significant reduction in the expression of cellular transcripts included in the DNA chip. This global approach to viral transcript analysis offers a powerful system for examining molecular transitions between lytic and latent virus infections associated with disease pathogenesis.

摘要

鼠γ疱疹病毒68基因组编码73个蛋白质编码开放阅读框,与人类γ(2)疱疹病毒有广泛的相似性,还有独特的基因和细胞同源物。我们使用基于寡核苷酸的微阵列对允许感染期间阶段特异性病毒RNA进行了转录组分析。使用这种方法,基于放线菌酮处理,M4、K3、ORF38、ORF50、ORF57和ORF73被指定为立即早期基因。微阵列分析还鉴定出10个具有早期表达动力学的转录本、32个具有早晚期表达动力学的转录本和29个具有晚期表达动力学的转录本。后一组主要由结构蛋白组成,相对于其他病毒转录本显示出高表达水平。此外,我们在放线菌酮和膦甲酸存在的情况下检测到了所有8种tRNA样转录本。用MHV-68进行裂解感染也导致DNA芯片中包含的细胞转录本表达显著降低。这种对病毒转录本分析的整体方法为研究与疾病发病机制相关的裂解性和潜伏性病毒感染之间的分子转变提供了一个强大的系统。

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