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从人胎儿组织RNA构建克隆文库。

Construction of cloned libraries from RNA of human fetal tissues.

作者信息

Kurnit D M, Wentworth B M, De Long L, Villa-Komaroff L

出版信息

Cytogenet Cell Genet. 1982;34(3):193-203. doi: 10.1159/000131807.

Abstract

We have constructed libraries of recombinant DNA plasmids containing sequences complementary to polyadenylated RNA from a variety of human midtrimester fetal tissues. The bacterial colonies containing these plasmids have been grown and replicated on nitrocellulose filters in a manner that facilitates permanent storage, rapid screening, and transportability to other laboratories. We screened a portion of the library for the presence of repetitive sequences and found that approximately 20% of the clones contain repetitive sequences. We have also shown that some clones contain nonrepetitive sequences. Pools of recombinant cDNA-containing plasmids devoid of repetitive sequences have been constructed to permit the chromosomal localization of a variety of actively transcribed sequences. The construction of such large, tissue-specific clone banks should facilitate the direct isolation, mapping, and characterization of normal and abnormal human genes.

摘要

我们构建了重组DNA质粒文库,其中包含与来自各种人类孕中期胎儿组织的聚腺苷酸化RNA互补的序列。含有这些质粒的细菌菌落已在硝酸纤维素滤膜上生长并复制,这种方式便于永久保存、快速筛选以及运输到其他实验室。我们对文库的一部分进行了重复序列筛选,发现约20%的克隆含有重复序列。我们还表明一些克隆含有非重复序列。已构建不含重复序列的含重组cDNA质粒池,以确定各种活跃转录序列的染色体定位。构建如此大规模的、组织特异性的克隆库应有助于直接分离、定位和鉴定正常和异常的人类基因。

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