Disruption of Thermus thermophilus genes by homologous recombination using a thermostable kanamycin-resistant marker
Disruption of Thermus thermophilus genes by homologous recombination using a thermostable kanamycin-resistant marker
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DOI：
10.1016/s0014-5793(01)02926-x
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发表时间：
2001-10-12
期刊：
影响因子：
3.5
通讯作者：
Kagamiyama, H
中科院分区：
文献类型：
--
作者：
Hashimoto, Y;Yano, T;Kagamiyama, H
一种极端嗜热细菌，嗜热栖热菌，基因被破坏的同源重组使用最近开发的，耐热卡那霉素抗性标记。首先，trpE基因被各种DNA结构破坏。当标记基因侧翼的同源区域的长度增加到超过最小长度（200-300 bp）时，转化效率呈指数增加。然后我们破坏了核苷酸切除修复系统的五个基因，并检查了它们的表型。这种方法的方便性和高可靠性，应促进其应用于高通量系统破坏这种嗜热细菌的基因。(C)2001年欧洲生物化学学会联合会。由Elsevier Science B. V.出版，版权所有。
Genes of an extremely thermophilic bacterium, Thermus thermophilus, were disrupted by homologous recombination using a recently developed, thermostable kanamycin-resistant marker. First, the trpE gene was disrupted with various constructions of DNA. The transformation efficiency was exponentially increased as the length of the homologous regions flanking the marker gene increased above the minimum length (200-300 bp). We then disrupted five genes of the nucleotide excision repair system and examined their phenotypes. The convenience and high reliability of this method should prompt its application to the high-throughput systematic disruption of the genes of this thermophilic bacterium. (C) 2001 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.