A thiostrepton resistance gene and its mutants serve as selectable markers in Geobacillus kaustophilus HTA426

Bioscience, Biotechnology, and Biochemistry
Keisuke WadaHirokazu Suzuki

Abstract

Effective utilization of microbes often requires complex genetic modification using multiple antibiotic resistance markers. Because a few markers have been used in Geobacillus spp., the present study was designed to identify a new marker for these thermophiles. We explored antibiotic resistance genes functional in Geobacillus kaustophilus HTA426 and identified a thiostrepton resistance gene (tsr) effective at 50 °C. The tsr gene was further used to generate the mutant tsr(H258Y) functional at 55 °C. Higher functional temperature of the mutant was attributable to the increase in thermostability of the gene product because recombinant protein produced from tsr(H258Y) was more thermostable than that from tsr. In fact, the tsr(H258Y) gene served as a selectable marker for plasmid transformation of G. kaustophilus. This new marker could facilitate complex genetic modification of G. kaustophilus and potentially other Geobacillus spp.

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Mar 19, 2015·Applied Microbiology and Biotechnology·Jyumpei KobayashiHirokazu Suzuki

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Citations

Jun 4, 2016·Genome Announcements·Katsumi DoiSeiya Ogata
Jan 20, 2018·The Journal of Antibiotics·Hirokazu SuzukiTakashi Ohshiro
Oct 13, 2018·Applied Microbiology and Biotechnology·Hirokazu Suzuki
Aug 19, 2018·Microbial Cell Factories·Megumi MiyanoKen-Ichi Yoshida
Oct 27, 2015·Genome Announcements·Kengo SakiharaKatsumi Doi
May 12, 2018·Microorganisms·Eivind B DrejerTrygve Brautaset
Apr 13, 2021·Frontiers in Microbiology·Hirokazu SuzukiTakashi Ohshiro

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Methods Mentioned

BETA
genetic modification
genetic modifications
electrophoresis
X-ray

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