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大鼠核糖体DNA克隆的分离与鉴定

Isolation and characterization of rat ribosomal DNA clones.

作者信息

Rothblum L I, Parker D L, Cassidy B

出版信息

Gene. 1982 Jan;17(1):75-7. doi: 10.1016/0378-1119(82)90102-0.

Abstract

Four EcoRI fragments, which contain the transcribed portion of the rat rDNA repeat, have been isolated from a rat genome library cloned in lambda Charon 4A vector. Three of the fragments, 9.6, 6.7, and 4.5 kb, from clones lambda ChR-B4, lambda Nr-42, and lambda ChR-C4B9, contained part of the 5'-NTS, the 5'-ETS, 18S rDNA, ITS-1, 5.8S rDNA, 28S rDNA and approximately 3.5 kb of the 3'-NTS. Two EcoRI fragments, from clones lambda ChR-B4 and lambda ChR-B7E12, which coded for the 5'-NTS, the ETS, and most of the 18S rDNA, differed by 1 kb near the EcoRI site upstream of the 5' terminus of 18S rRNA. Restriction maps of the cloned DNA fragments were constructed by cleavage of the fragments with various restriction endonucleases and Southern hybridization with 18S, 5.8S, and 28S rRNA. These maps were confirmed and extended by subcloning several regions of the repeat in pBR322.

摘要

从克隆于λ噬菌体Charon 4A载体的大鼠基因组文库中分离出了四个包含大鼠rDNA重复序列转录部分的EcoRI片段。来自克隆λChR - B4、λNr - 42和λChR - C4B9的三个片段,大小分别为9.6、6.7和4.5 kb,包含5'-NTS、5'-ETS、18S rDNA、ITS - 1、5.8S rDNA、28S rDNA以及约3.5 kb的3'-NTS的部分序列。来自克隆λChR - B4和λChR - B7E12的两个编码5'-NTS、ETS和大部分18S rDNA的EcoRI片段,在18S rRNA 5'末端上游的EcoRI位点附近相差1 kb。通过用各种限制性内切酶切割片段并与18S、5.8S和28S rRNA进行Southern杂交,构建了克隆DNA片段的限制性图谱。通过将重复序列的几个区域亚克隆到pBR322中,对这些图谱进行了确认和扩展。

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