Characterizing and Genetically Modifying Oscillatoria to Degrade BPA

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Long, Grace

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Bisphenol A (BPA) is becoming an increasing environmental concern. It is a hormone disruptor, and its presence in natural waterways causes some aquatic species to experience problems in growth, reproduction, and development. An effective BPA removal system is needed. Genetically modifying a type of cyanobacteria—Oscillatoria—with a gene from a fungus—Trametes versicolor—may provide a solution. The T. versicolor’s gene produces a laccase with the ability to degrade BPA in the presence of butylhydroxytoluene—a molecule naturally produced by Oscillatoria. Before transforming Oscillatoria, however, it is necessary to understand the genus. In this study, I characterized the growth of Oscillatoria brevis, Oscillatoria lutea, and Oscillatoria prolifera as well as their ability to naturally fluoresce. Additionally, I determined some of the species’ antibiotic resistances: O. brevis was found to have some resistance to carbenicillin, gentamicin, and kanamycin; O. lutea was found to have some resistance to carbenicillin, gentamicin, and kanamycin; and O. prolifera was found to have some resistance to gentamicin, kanamycin, and spectinomycin. Furthermore, I attempted to conjugate the Oscillatoriae with a plasmid containing GFP and antibiotic resistance to carbenicillin, kanamycin, or spectinomycin. Because the cyanobacteria are not axenic (pure, without contaminants), different organisms in the microbial community were unintentionally transformed instead. Finally, I also created genetic parts to be used in Golden Gate Assembly once the conjugation procedure is successful.



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