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iGEM CINVESTAV_IPN-UNAM

The PrrA dependent promoter was used in purple non-sulfur photosynthetic bacteria (PNSP) R. sphaeroides and R. palustris, using different conditions. For test the promoter we used as a reporter GFP.

Fig 1. Construction of PrrA dependent promoter.

We employed flow cytometery for mesuring the expression of GFP. The conditions we used for test the PrrA dependent promoter were: aerobic/darness, anaerobic/light, anaerobic/darknness. The results were as follows.

R. sphaeroides

Graphic 1.: Quantitative analysis of bacterial population with GFP expression (GFP+), under above condition, using flow cytometry (Attune cytometer Applied Biosystems), background signals were eliminated using a negative control.

Graphic 2. Median fluorescence Intensity of the conjugated R. sphaeroides strain. Analysis of fluorescence in bacterial populations (1000 bacteria), under above conditions, using flow cytometry (Attune cytometer Applied Biosystems), background signals were eliminated using a negative control.

R. palustris

Graphic 3.: Quantitative analysis of bacterial population with GFP expression (GFP+), under above condition, using flow cytometry (Attune cytometer Applied Biosystems), background signals were eliminated using a negative control.

Graphic 4. Median fluorescence Intensity of the conjugated R. palustris strain. Analysis of fluorescence in bacterial populations (1000 bacteria), under above conditions, using flow cytometry (Attune cytometer Applied Biosystems), background signals were eliminated using a negative control.

Results above shows that the PrrA dependent promoter is functional in our two PNSP bacteria.

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