DIFFERENTIAL DISPLAY OF EUKARYOTIC MESSENGER-RNA BY MEANS OF THE POLYMERASE CHAIN-REACTION
DIFFERENTIAL DISPLAY OF EUKARYOTIC MESSENGER-RNA BY MEANS OF THE POLYMERASE CHAIN-REACTION
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DOI：
10.1126/science.1354393
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发表时间：
1992-08-14
期刊：
影响因子：
56.9
通讯作者：
PARDEE, AB
中科院分区：
文献类型：
--
作者：
LIANG, P;PARDEE, AB
需要有效的方法来识别和分离那些在不同细胞中或在不同条件下差异表达的基因。本文介绍了一种利用聚合酶链式反应分离和克隆单个信使RNA的方法。关键是使用一组寡核苷酸引物，一个锚定在mRNAs子集的多聚腺苷酸尾上，另一个短而任意的序列，以便它在相对于第一个引物的不同位置进行退火。经逆转录后扩增，DNA测序凝胶鉴定。当使用多个引物组时，获得的扩增互补DNA片段的重复性模式显示出对任一引物序列特异性的强烈依赖性。
Effective methods are needed to identify and isolate those genes that are differentially expressed in various cells or under altered conditions. This report describes a method to separate and clone individual messenger RNAs (mRNAs) by means of the polymerase chain reaction. The key element is to use a set of oligonucleotide primers, one being anchored to the polyadenylate tail of a subset of mRNAs, the other being short and arbitrary in sequence so that it anneals at different positions relative to the first primer. The mRNA subpopulations defined by these primer pairs were amplified after reverse transcription and resolved on a DNA sequencing gel. When multiple primer sets were used, reproducible patterns of amplified complementary DNA fragments were obtained that showed strong dependence on sequence specificity of either primer.