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Disruption of Thermus thermophilus genes by homologous recombination using a thermostable kanamycin‐resistant marker
Author(s) -
Hashimoto Yasuko,
Yano Takato,
Kuramitsu Seiki,
Kagamiyama Hiroyuki
Publication year - 2001
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(01)02926-x
Subject(s) - thermus thermophilus , homologous recombination , gene , biology , genetics , thermus , transformation (genetics) , thermophile , homologous chromosome , kanamycin , plasmid , recombination , genetic recombination , microbiology and biotechnology , bacteria , escherichia coli
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.