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Violacein is a purple-pigmented small metabolite that has significant biomedical and ecological importance. Significantly, violacein has been studied for its killing effect against Batrachochytrium dendrobatidis (Bd), a toxic and invasive chytrid fungus causing dramatic declines of amphibians worldwide. Violacein production in bacteria is completed through a five gene operon (vioA-E). Violacein-producing species are of scientific interest due to their potential use as an environmentally-friendly bioremediation agent against the invasive Bd fungus. Here we report the development of the genome editing method, CRISPR-Cas9, to disrupt vioA of a Hudson Valley native Iodobacter species, BJB302. By utilizing CRISPR-Cas9 mechanisms and comparative genomic strategies, we offer insight into violacein biosynthesis and regulation. Our work is also one of the first to contribute to CRISPR-Cas9 genome editing in undergraduate research. By comparing the violacein biosynthesis operon among several different violacein-producing strains, we have furthered the understanding of the violacein metabolite and its biosynthesis. Thus, this study provides insight into improving violacein-producing strains as a bioremediation agent.
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Mitchell, Cailey Marie, "Employing CRISPR-Cas9 to Disrupt the Violacein Biosynthesis Operon of BJB302, a Unique Violacein-Producing Isolate from the Hudson Valley" (2021). Senior Projects Spring 2021. 22.
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