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利用聚合酶链反应检测潜伏感染和活跃感染小鼠神经节中单纯疱疹病毒1型基因的表达。

Detection of herpes simplex virus type 1 gene expression in latently and productively infected mouse ganglia using the polymerase chain reaction.

作者信息

Lynas C, Laycock K A, Cook S D, Hill T J, Blyth W A, Maitland N J

机构信息

Department of Ophthalmology, Medical School, Bristol, U.K.

出版信息

J Gen Virol. 1989 Sep;70 ( Pt 9):2345-55. doi: 10.1099/0022-1317-70-9-2345.

Abstract

The polymerase chain reaction (PCR) was employed to detect herpes simplex virus (HSV) sequences in the DNA, and HSV gene expression in total cell RNA, extracted from cervical and trigeminal ganglia of mice during productive and latent infection with HSV-1, strain SC16. Such gene expression was detected in 1 microgram or less of RNA, the quantity anticipated to be present in one or two cervical ganglia. Within the limits of the primers available, gene expression during latency appeared to be restricted to the latency-associated transcript (LAT). The 195 base portion of the LAT amplified by the PCR was sequenced and found to contain several base changes and deletions with respect to published sequences for different HSV strains. These mutations, within the putative open reading frame 2 of the LAT, formed stop or terminator signals, which suggests that the LAT does not act to establish or maintain latency through translation to a protein. The primers for the LAT also amplified a 300 bp fragment from any murine and some other mammalian RNAs. Apart from the oligonucleotide primers, this fragment did not show any homology with HSV.

摘要

采用聚合酶链反应(PCR)检测DNA中的单纯疱疹病毒(HSV)序列,以及从感染1型单纯疱疹病毒(HSV-1)SC16株处于增殖性感染和潜伏感染状态的小鼠颈神经节和三叉神经节中提取的总细胞RNA中的HSV基因表达。在1微克或更少的RNA中检测到了这种基因表达,预计一两个颈神经节中会存在这样的RNA量。在所使用引物的限度内,潜伏期间的基因表达似乎仅限于潜伏相关转录本(LAT)。对通过PCR扩增的LAT的195个碱基部分进行测序,发现相对于不同HSV毒株的已发表序列,其包含多个碱基变化和缺失。这些位于LAT推定开放阅读框2内的突变形成了终止信号,这表明LAT并非通过翻译为蛋白质来建立或维持潜伏状态。用于LAT的引物还从任何小鼠和一些其他哺乳动物的RNA中扩增出一个300 bp的片段。除了寡核苷酸引物外,该片段与HSV没有任何同源性。

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