Isola N R, Harn H J, Cooper D L
University of Pittsburgh School of Medicine, Department of Pathology, PA 15261.
Biotechniques. 1991 Nov;11(5):580, 582.
We describe an expeditious method for the isolation of cDNA clones utilizing PCR-based amplification of target sequences from cDNA libraries. This method is rapid, less labor-intensive and inexpensive when compared with screening libraries with radiolabeled probes. This method can be applied to isolate multiple members of a protein family as well as homologous genes in different species by designing appropriate primers to amplify the most conserved regions. Utilizing this method, a novel reticulocyte CD44 transcript was isolated.
我们描述了一种利用基于PCR的从cDNA文库中扩增目标序列来分离cDNA克隆的快速方法。与用放射性标记探针筛选文库相比，该方法快速、劳动强度低且成本低廉。通过设计合适的引物扩增最保守区域，此方法可用于分离蛋白质家族的多个成员以及不同物种中的同源基因。利用该方法，分离出了一种新型的网织红细胞CD44转录本。
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