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用于直接筛选与低丰度哺乳动物细胞mRNA相对应的cDNA克隆的载体。

Vectors for the direct selection of cDNA clones corresponding to mammalian cell mRNA of low abundance.

作者信息

Kowalski J, Smith J H, Ng N, Denhardt D T

出版信息

Gene. 1985;35(1-2):45-54. doi: 10.1016/0378-1119(85)90156-8.

Abstract

We have constructed two cDNA cloning vectors which (1) carry the intergenic region of phage f1 and (2) permit efficient cloning (by the Okayama-Berg procedure) of full-length copies of mammalian mRNA in either orientation. Infection of cells harboring these vectors with f1 phage results in the encapsidation of single-stranded (ss) plasmid DNA carrying either the sense or the anti-sense sequence of the cDNA inserts. The complementary nature of the cDNA inserts in two such cDNA libraries facilitates preparative hybridization procedures. These vectors have general applicability to any eukaryotic system where changes in the abundance of mRNA transcripts are to be measured and the corresponding cDNA clones isolated.

摘要

我们构建了两种cDNA克隆载体,它们(1)携带噬菌体f1的基因间隔区,(2)允许通过冈本-伯格方法高效克隆哺乳动物mRNA的全长拷贝,且两种方向均可。用f1噬菌体感染携带这些载体的细胞,会导致携带cDNA插入片段的正义或反义序列的单链(ss)质粒DNA被包装。两个这样的cDNA文库中cDNA插入片段的互补性质便于制备性杂交程序。这些载体普遍适用于任何要测量mRNA转录本丰度变化并分离相应cDNA克隆的真核系统。

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