New strategies for isolation of low abundance viral and host cDNAs: application to cloning of the hepatitis E virus and analysis of tissue-specific transcription.
New strategies for isolation of low abundance viral and host cDNAs: application to cloning of the hepatitis E virus and analysis of tissue-specific transcription.
复制标题
分离低丰度病毒和宿主 cDNA 的新策略：应用于戊型肝炎病毒克隆和组织特异性转录分析。
DOI：
10.1055/s-2008-1040398
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发表时间：
1992
影响因子：
4.2
通讯作者：
Lovett,M
中科院分区：
文献类型：
--
作者：
Reyes,GR;Bradley,DW;Lovett,M
Any strategy for the screening and cloning of cDNAs must cope with the dual problems of cDNA sequence complexity and abundance. This is true whether one is searching for a viral RNA or DNA genome, present in a small percentage of the target tissue, or if one is searching for a low abundance host transcript from the same type of source material. Unfortunately, it is frequently the case that the very cDNAs we would most like to isolate, those that encode a previously uncharacterized viral genome or perhaps the regulators of viral or host gene expression and cellular growth, are either transiently expressed or expressed at low levels in subsets of cells within a given tissue. The target molecules, be they viral or cellular in origin, therefore represent a minor component in a complex mixture of transcripts making the identification of low abundance cDNAs the molecular equivalent of searching for a" needle in a haystack." In this article we describe strategies that we have developed to isolate rapidly these low abundance cDNAs. These strategies have particular application to the identification of new viral entities and in the isolation of genetic loci where the level of expression precludes the application of more traditional techniques. We illustrate these new methods by the molecular cloning and isolation of the hepatitis E virus and also describe their application to the analysis of tissue-specific transcription.
