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洛里斯2,一种黏粒,带有转录终止子,可使载体基因免受插入片段内启动子的干扰:对DNA产量和克隆插入片段频率的影响。

Lorist2, a cosmid with transcriptional terminators insulating vector genes from interference by promoters within the insert: effect on DNA yield and cloned insert frequency.

作者信息

Gibson T J, Coulson A R, Sulston J E, Little P F

出版信息

Gene. 1987;53(2-3):275-81. doi: 10.1016/0378-1119(87)90016-3.

Abstract

Transcription terminators have been included in a phage-lambda-replicon-based cosmid vector, Lorist2, to insulate vector genes against transcriptional interference from cloned insert DNA. DNA yields of recombinant clones containing Escherichia coli genomic DNA inserts are more even for Lorist2 than with its progenitor LoristB. However, the terminators provide only a partial reduction in the over-representation of r X DNA-containing clones generally observed in cosmid libraries of Caenorhabditis elegans DNA, suggesting that causes other than transcriptional readthrough into the vector contribute to this problem.

摘要

转录终止子已被包含在基于噬菌体λ复制子的黏粒载体Lorist2中,以隔离载体基因,防止其受到来自克隆插入DNA的转录干扰。对于含有大肠杆菌基因组DNA插入片段的重组克隆,Lorist2的DNA产量比其亲本载体LoristB更为均匀。然而,这些终止子只能部分减少在秀丽隐杆线虫DNA黏粒文库中普遍观察到的含rX DNA克隆的过度富集现象,这表明除了转录通读进入载体之外,还有其他原因导致了这个问题。

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