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Identification Of Genes Involved In The Production Of Novel Antimicrobial Products Capable Of Inhibiting Multi-Drug Resistant Pathogens

Harris, Ryan A
http://rave.ohiolink.edu/etdc/view?acc_num=bgsu1562068774992706

Abstract Details

2019, Master of Science (MS), Bowling Green State University, Biological Sciences.
The research described here focuses on the phylogenetic characterization of water-derived pseudomonads and their antagonistic activity against multi-drug resistance (MDR) P. aeruginosa and Burkholderia species. Phylogenetic work was based on the gyrB housekeeping gene. Genetic techniques have been optimized and employed to identify genes associated with antimicrobial production via transposon (Tn) mutagenesis using a triparental mating system approach with Pseudomonas as the model organism. This study expands on theses previous studies in the lab to identify biosynthetic gene clusters (BGC) involved in production of novel antibiotics capable of inhibiting the growth of MDR pathogens. We utilize a previously optimized workflow to identify genes from environmental isolates involved in the inhibition of MDR P. aeruginosa and species within the Burkholderia cepacia complex (Bcc). We show that both MDR Bcc and P. aeruginosa pathogens were inhibit by environmental Pseudomonas strains. Out of 7,784 interactions, 210 of these were antagonistic. Superkillers (SK), defined as strains that inhibit ≥3 of MDR pathogens, were selected for optimization of Tn mutagenesis to identify gene cluster whose products inhibit these MDR pathogens. Only six out of the 24 SK’s were capable of this process. Out of these six, three were selected for large scale mutagenesis to identify loss of inhibition (LOI) mutants. Four LOI mutations were found for strain S5F11, one of which had an insertion within a BGC predicted to produce an NRPS complex. Seven LOI mutants were found for S3E7. Although none of these insertions were identified within a BGC, genes have been identified that are observed to be heavily involved in antibiotic production. This study suggests that environmental Pseudomonas strains have the capacity to inhibit the growth of CF-derived MDR pathogens. Using Tn mutagenesis, we have identified novel loci that are associated with antibiotic production.
Hans Wildschutte, PhD (Advisor)
Timothy Davis, PhD (Committee Member)
Robert Huber, PhD (Committee Member)
91 p.
Biology
Microbiology; Antibiotic resistance; Pseudomonas; Pseudomonas aeruginosa; Antibiotics; Transposon mutagenesis; Arbitrary PCR; Antibiotic discovery; Novel antibiotics; Cystic fibrosis; multi-drug resistance; Biosynthetic gene clusters; Burkholderia

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Citations

  • Harris, R. A. (2019). Identification Of Genes Involved In The Production Of Novel Antimicrobial Products Capable Of Inhibiting Multi-Drug Resistant Pathogens [Master's thesis, Bowling Green State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=bgsu1562068774992706

    APA Style (7th edition)

  • Harris, Ryan. Identification Of Genes Involved In The Production Of Novel Antimicrobial Products Capable Of Inhibiting Multi-Drug Resistant Pathogens. 2019. Bowling Green State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=bgsu1562068774992706.

    MLA Style (8th edition)

  • Harris, Ryan. "Identification Of Genes Involved In The Production Of Novel Antimicrobial Products Capable Of Inhibiting Multi-Drug Resistant Pathogens." Master's thesis, Bowling Green State University, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=bgsu1562068774992706

    Chicago Manual of Style (17th edition)

bgsu1562068774992706
436
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This open access ETD is published by Bowling Green State University and OhioLINK.