Part:BBa_K4117666:Experience

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Applications of BBa_K4117666

We added the GFP downstream of the RNA thermometer (RBS) to characterize the efficiency of RNA thermometer.

Fig.1 RNA thermometer verification pathway

In order to verify the function of RNA thermometers, We cultured the strain at 37°C overnight, and then aliquots, adding pre-prepared flasks filled with 37°C and 28°C LB medium to make the final volume 20ml, then measured the initial OD600 and GFP fluorescence intensity. Samples were taken once every half-hour, OD600 and GFP fluorescence intensity were measured.

GFP measurement results are shown in Fig.2.

Fig.2 GFP fluorescence intensity after incubating in different temperatures

And we used a fluorescence microscope for observation. The bacterial liquid under the fluorescence microscope is as Fig. 3:

Fig.4 Images under fluorescene microscope (A) No GFP in engineerd bacteria incubated at 28℃ (B) Bright field image of bacteria incubated at 28℃ (C) The field of view after superposition of A and B (D) GFP in engineerd bacteria incubated at 37℃ (E) Bright field image of bacteria incubated at 37℃ (F) The field of view after superposition of D and E (fluorescene microscope 10 x 100)

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