Experimental Results

Testing of Or as a Promoter

The backbone of the radiation sensor in our project relies on the operating region Or, and the two promoters one on each end of Or. In order to test the function ability of the of the pr promoter we attached a part called mcherry, which has rfp to the promoter. We used the TECAN in our lab to measure the fluorescence of our samples over time. The data collected while the samples were in exponential growth was used to determine a rate of expression of the rfp. Compared to the control, the rate of fluorescence was much higher in our test subject, about 4227 compared to 418 (fu/OD600nm). This suggests that the promoter on Or is functional.

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Collaboration

Throughout the summer we connected with other iGEM teams interested in radiation-related projects. The NYC Wetware team in particular was studying the effects of genes cloned from the organism Deinococcus radiodurans on radiation resistance in E.coli. We were given the honor of helping them characterize one of their strains TS TR, a strain capable of producing the molecule trypanothione.

Using the gamma facilities provided by the Radiation Science and Engineering Center at Penn state we were able to carry out tests of gamma irradiation of their strain and subsequently measure cell viability.

Aliquots of culture (5ml each) inoculated with the TS TR strain were added to tubes which were irradiated by a cobalt-60 gamma source at a dose rate of 25 gys/min for total doses ranging from 0-800 gys. After irradiation 0.02 ml of the irradiated sample was inoculated into 5 ml of SOB media. The OD₆₀₀ was measured 12 hours later. The results are shown below.