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Cloning and Sequencing of Mercury Resistance Operons from the Enterobacter sp. YSU and Stenotrophomonas maltophilia OR02

Muhindi, Stephen W
http://rave.ohiolink.edu/etdc/view?acc_num=ysu1525449851977263

Abstract Details

2018, Master of Science in Biological Sciences, Youngstown State University, Department of Biological Sciences and Chemistry.
Enterobacter sp. YSU and Stenotrophomonas maltophilia OR02 (S. maltophilia 02) were isolated from a heavy-metal contaminated site in Oak Ridge, TN. Both strains are resistant to salts of copper, cadmium, zinc, selenium, arsenic, gold and mercury. Known primers for the mercury resistance genes, merR and merP, were used to PCR amplify, clone and sequence DNA segments from each strain. Then, Basic Local Alignment Search Tool (BLAST) analysis of the sequences identified reference sequences used to sequence the rest of the mer genes in each strain. These reference sequences contained the genes, merRTPCADE, for Enterobacter sp. YSU and the genes, merRTPADE, for S. maltophilia 02. Alignments using other related merR and merE genes were used to design PCR primer to amplify both mer operons which were cloned into the plasmid, pSC-A-amp/kan, from the StrataClone PCR Cloning Kit. Liquid culture experiments in LB broth showed that the minimal inhibitory concentrations (MICs) were 150 µM and 190 µM HgCl2, for the Enterobacter sp. YSU and S. maltophilia 02 cloned fragments, respectively, and 30 µM HgCl2 for the vector control. The bioinformatics tool, Molecular Evolutionary Genetic Analysis (MEGA 7), generated a phylogenetic tree that showed that the MerA protein from Enterobacter sp. YSU is related to other Enterobacter MerA proteins and that the MerA protein from S. maltophilia 02 is related to other Stenotrophomonas MerA proteins.
Jonathan Caguiat, PhD (Advisor)
David Asch, PhD (Committee Member)
Xiangjia Min, PhD (Committee Member)
35 p.
Bioinformatics; Biology; Cellular Biology; Microbiology; Molecular Biology
Mercury resistant genes, Mercuric ion, Enterobacter, Stenotrophomonas

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Citations

  • Muhindi, S. W. (2018). Cloning and Sequencing of Mercury Resistance Operons from the Enterobacter sp. YSU and Stenotrophomonas maltophilia OR02 [Master's thesis, Youngstown State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1525449851977263

    APA Style (7th edition)

  • Muhindi, Stephen. Cloning and Sequencing of Mercury Resistance Operons from the Enterobacter sp. YSU and Stenotrophomonas maltophilia OR02. 2018. Youngstown State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ysu1525449851977263.

    MLA Style (8th edition)

  • Muhindi, Stephen. "Cloning and Sequencing of Mercury Resistance Operons from the Enterobacter sp. YSU and Stenotrophomonas maltophilia OR02." Master's thesis, Youngstown State University, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1525449851977263

    Chicago Manual of Style (17th edition)

ysu1525449851977263
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© 2018, all rights reserved.
This open access ETD is published by Youngstown State University and OhioLINK.
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