Development of qPCR Based Method to Quantify Colonization of an Engineered Bacterium in Gut

dc.contributor.author McMullen, Alexandra (Alex)
dc.contributor.department Biomedical Sciences
dc.contributor.majorProfessor Anumantha Kanthasamy
dc.date 2020-03-02T19:44:44.000
dc.date.accessioned 2020-06-30T01:35:10Z
dc.date.available 2020-06-30T01:35:10Z
dc.date.copyright Tue Jan 01 00:00:00 UTC 2019
dc.date.issued 2019-01-01
dc.description.abstract <p>Parkinson’s Disease is the leading neurodegenerative disease in the western world, caused by degeneration of neurons in the substantia nigra. This causes a decrease of dopamine in nigrostriatal neural circuitry, leading to motor and nonmotor symptoms which can be incredibly debilitating for the patient. Current dopamine supplementation therapy does not provide a steady state neurotransmitter level and can lead to dyskinesias and dystonia’s. However, <em>Escherichia coli Nissle </em>(EcN) bacteria have been shown to prove effective therapeutic treatments that can be engineered to specifically treat certain diseases, such as Phenylketonuria. EcN can be engineered to upregulate production of L-DOPA by adding an extra <em>HpaBC </em>gene. Administration of L-DOPA through ECN can maintain steady state dopamine level and thereby normalizing dopaminergic neural pathways for patients suffering from Parkinson’s Disease. The goal of this study was to monitor EcN-DOPA gut colonization in mouse models of Parkinson’s disease using a qPCR method. We generated a unique primer set for ECN-L-DOPA and determined its utility in measuring the colonization. Our results showed that ECN-L-DOPA primer effectively amplifies the gene and detected the engineered bacteria in the fecal sample. Further evaluation in Mitopark transgenic mouse model of Parkinson’s disease, qPCR method successfully detected colonization profile of ECN-DOPA and correlated with the plasma level with plasma L-DOPA level. Collectively, our results show qPCR method is very amenable for monitoring gut colonization in microbiome-based therapy.</p>
dc.format.mimetype Word
dc.identifier archive/lib.dr.iastate.edu/creativecomponents/457/
dc.identifier.articleid 1295
dc.identifier.contextkey 14361421
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath creativecomponents/457
dc.identifier.uri https://dr.lib.iastate.edu/handle/20.500.12876/17019
dc.source.bitstream archive/lib.dr.iastate.edu/creativecomponents/457/McMullen_Final_Creative_Component_For_Submission.docx|||Sat Jan 15 00:21:31 UTC 2022
dc.source.bitstream archive/lib.dr.iastate.edu/creativecomponents/457/auto_convert.pdf|||Sat Jan 15 00:21:33 UTC 2022
dc.subject.disciplines Biological Phenomena, Cell Phenomena, and Immunity
dc.subject.disciplines Digestive, Oral, and Skin Physiology
dc.subject.disciplines Digestive System Diseases
dc.subject.disciplines Nervous System Diseases
dc.subject.keywords Parkinson's Disease
dc.subject.keywords EcN
dc.title Development of qPCR Based Method to Quantify Colonization of an Engineered Bacterium in Gut
dc.type article
dc.type.genre creativecomponent
dspace.entity.type Publication
relation.isOrgUnitOfPublication 184db3f2-d93f-4571-8ad7-07c8a9e6a5c9
thesis.degree.discipline Biomedical Sciences
thesis.degree.level creativecomponent
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