Team:Austin UTexas/Results


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Figure 4: amilCP expressed in E. coli and in liquid LB. Credit: Riya Sreenivasan

GOX Sequences as Putative Promoters

Three endogenous upstream regions of loci on the Gluconobacter oxydans chromosome were reported to show increased mRNA synthesis as pH decreased, were isolated and obtained, as seen in table 1 (Hanke, et al., 2012). Using Golden Gate assembly, these putative promoters have been placed on the Golden Gate entry vector pYTK001 for later use. By utilizing these pH-sensitive promoters with different reporters and transforming them into multiple organisms in kombucha, the visualization of the microbes and their location in kombucha would be possible (Lee, et al., 2015). This will serve as a stepping stone into further understanding how the microbiome of kombucha changes as it brews as well as determining organism concentration specific times during the brewing process.

Figure 5. Spun down P-atp2 constructs compared to controls in pH6-9. There is no clear gradient change in color expression. Credit: Ian Overman and Alex Alario

Table 1:The Three Endogenous GOX Sequences
Locus Tag Predicted Functions mRNA ratio pH4/pH6
GOX0647 Putative exporter protein, ArAE family 12.91
GOX0890 Hypothetical protein GOX0890 4.93
GOX1841 Hypothetical protein GOX1841 3.36

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