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Identification of a putative metK selenite resistance gene in Stenotrophomonas maltophilia OR02

Marinelli, Zachary A
http://rave.ohiolink.edu/etdc/view?acc_num=ysu1516345597337997

Abstract Details

2017, Master of Science in Biological Sciences, Youngstown State University, Department of Biological Sciences and Chemistry.
Stenotrophomonas maltophilia OR02 (S02) is a multi-metal resistant strain that was isolated from a metal-contaminated site in Oak Ridge, TN. It grows in the presence of 20 mM sodium selenite and produces a red precipitate which is probably elemental selenium, and a stale garlic odor, which is probably methyl-selenide. The reduction of selenite to selenide may be dependent on the role of a glutathione reductase. Then, methyl-selenide may be produced by a thiopurine methyltransferase. A selenite-sensitive mutant was generated by introducing the EZ-Tn5 transposome into S02. This transposon, which carried a kanamycin resistance gene, randomly incorporated itself into the S02 genome and generated thousands of kanamycin resistant transformants. Replica plating of 880 transformants yielded one selenite-sensitive S02 mutant, AX55. Liquid culture experiments showed that the minimal inhibitory concentration (MIC) for AX55 was 10 mM selenite, whereas the MIC for the S02 was 30 mM. DNA sequencing and Basic Local Alignment Search Tool (BLAST) analysis showed that the transposon interrupted the 5’ untranslated region (UTR) of a putative metK gene. The metK gene encodes the protein S-adenosylmethionine (SAM) synthetase, which catalyzes the formation of SAM. These results confirm the requirement of SAM for selenite methylation, which may be catalyzed by a thiopurine methyltransferase. Furthermore, the decreased production of selenium nanoparticles by AX55 suggests the reduction mechanism is also dependent on SAM.
Jonathan Caguiat, PhD (Advisor)
David Asch, PhD (Committee Member)
Chester Cooper, PhD (Committee Member)
82 p.
Biology; Environmental Science; Genetics; Microbiology; Molecular Biology
Stenotrophomonas maltophilia; metK; S-adenosylmethionine; selenite resistance; selenite detoxification; bioremediation

Recommended Citations

Citations

  • Marinelli, Z. A. (2017). Identification of a putative metK selenite resistance gene in Stenotrophomonas maltophilia OR02 [Master's thesis, Youngstown State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1516345597337997

    APA Style (7th edition)

  • Marinelli, Zachary. Identification of a putative metK selenite resistance gene in Stenotrophomonas maltophilia OR02. 2017. Youngstown State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ysu1516345597337997.

    MLA Style (8th edition)

  • Marinelli, Zachary. "Identification of a putative metK selenite resistance gene in Stenotrophomonas maltophilia OR02." Master's thesis, Youngstown State University, 2017. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1516345597337997

    Chicago Manual of Style (17th edition)

ysu1516345597337997
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© 2017, all rights reserved.
This open access ETD is published by Youngstown State University and OhioLINK.