Disruption of Thermus thermophilus genes by homologous recombination using a thermostable kanamycin-resistant marker

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Abstract

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. © 2001 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.

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Hashimoto, Y., Yano, T., Kuramitsu, S., & Kagamiyama, H. (2001). Disruption of Thermus thermophilus genes by homologous recombination using a thermostable kanamycin-resistant marker. FEBS Letters, 506(3), 231–234. https://doi.org/10.1016/S0014-5793(01)02926-X

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