Demonstrate
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Kombucha Strains
- Successfully isolated microbes from various samples of kombucha.
- Identified strains of bacteria and yeast using rRNA gene sequencing.
- Characterized each of the isolated microbes to facilitate further experimentation.
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Recapitulation
- In a process called "recapitulation," we successfully created a kombucha-like culture by adding individual strains of microbes instead of a living culture containing the entire kombucha microbiome.
- Determined that the microbe Ga. hansenii is essential for the fermentation of kombucha.
- Determined that two distinct strains of the yeast Lachancea fermentati are necessary for the fermentation of kombucha, including one that appears to produce high quantities of C02.
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Conjugation
- Attempted conjugation with G. oxydans.
- Performed minimum inhibitory concentration experiments between G. oxydans and spectinomycin, carbenicillin and kanamycin.
- Determined that G. oxydans is resistant to spectinomycin and carbenicillin.
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Ethanol
- Found literature describing sequences for genes involved in the metabolism of ethanol to acetic acid in the bacterium Ga. hansenii.
- Designed Golden Gate parts for the assembly of these genes into a functional construct.
- Used a bromothymol blue assay to compare changes in pH resulting from fermentation in multiple strains of Lachancea fermentati isolated from our kombucha.
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pH Sensors
- Successfully created a neutral pH sensor with a reporter.
- Further characterized the P-atp2 Biobrick.
- Found literature describing three putative promoters in Gluconobacter oxydans that increase transcription under acidic conditions, and currently characterizing these sequences.
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