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| We were able to successfully measure YFP fluorescence intensity and fluorescence spectra of YFP-expressing <i>C. reinhardtii</i> clones in comparison to the wild type (WT). The results showed that our clone exhibited a higher fluorescence intensity at 528 nm than the WT (YFP emission peak) and the fluorescence spectrum of YFP confirmed the presence of the yellow fluorescent protein. This YFP-expressing clone also allowed us to characterize the light induction of the PsaD promoter by doing a time-resolved measurement of the fluorescence intensity. | | We were able to successfully measure YFP fluorescence intensity and fluorescence spectra of YFP-expressing <i>C. reinhardtii</i> clones in comparison to the wild type (WT). The results showed that our clone exhibited a higher fluorescence intensity at 528 nm than the WT (YFP emission peak) and the fluorescence spectrum of YFP confirmed the presence of the yellow fluorescent protein. This YFP-expressing clone also allowed us to characterize the light induction of the PsaD promoter by doing a time-resolved measurement of the fluorescence intensity. |
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| During this measurement we exposed <i>C. reinhardtii</i> cultures which contained our YFP construct to different growth conditions. One was exposed only to the dark, the other to synchronized growth conditions(10 hours dark, 14 hours illuminated). Then, we started a time-resolved fluorescence intensity measurement in the dark, with a WT control. After approximately four hours, we activated a light source and exposed the cultures to light, thus activating the light inducible PsaD promoter. Our results showed that for the dark and synchronized cultures containing the YFP construct a peak in fluorescence intensity could clearly be seen after the light induction. This proved the light induction of the PsaD promoter. If you are interested in this measurement, please visit the page of our YFP mVenus construct in the iGEM registry <a href="http://parts.igem.org/Part:BBa_K2984019">here</a>. | | During this measurement we exposed <i>C. reinhardtii</i> cultures which contained our YFP construct to different growth conditions. One was exposed only to the dark, the other to synchronized growth conditions(10 hours dark, 14 hours illuminated). Then, we started a time-resolved fluorescence intensity measurement in the dark, with a WT control. After approximately four hours, we activated a light source and exposed the cultures to light, thus activating the light inducible PsaD promoter. Our results showed that for the dark and synchronized cultures containing the YFP construct a peak in fluorescence intensity could clearly be seen after the light induction. This proved the light induction of the PsaD promoter. If you are interested in this measurement, please visit the page of our YFP mVenus construct in the iGEM registry <a href="http://parts.igem.org/Part:BBa_K2984019">here</a>. |
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| <img class="is-revealing" src="https://static.igem.org/mediawiki/2019/7/73/T--Humboldt_Berlin--YFP_fluorescence_intensity.png" alt="fluorescence intensity" /> | | <img class="is-revealing" src="https://static.igem.org/mediawiki/2019/7/73/T--Humboldt_Berlin--YFP_fluorescence_intensity.png" alt="fluorescence intensity" /> |
− | <caption>Fig. - Fluorescence intensity of <i>C. reinhardtii</i> WT and a YFP-carrying clone, in decreasing optical density of the cell culture. Excitation at 490 nm and emission measurement at 528 nm. The results clearly show that the fluorescence of the YFP-expressing clone is higher than the autofluorescence of the WT algae.</caption> | + | <caption>Fig. - Fluorescence intensity of C. reinhardtii WT and a YFP-carrying clone, in decreasing optical density of the cell culture. Excitation at 490 nm and emission measurement at 528 nm. The results clearly show that the fluorescence of the YFP-expressing clone is higher than the autofluorescence of the WT algae.</caption> |
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| <img class="is-revealing" src="https://static.igem.org/mediawiki/2019/5/50/T--Humboldt_Berlin--YFP_fluorescence_difference_spectrum.png" alt="fluorescence difference spectrum" /> | | <img class="is-revealing" src="https://static.igem.org/mediawiki/2019/5/50/T--Humboldt_Berlin--YFP_fluorescence_difference_spectrum.png" alt="fluorescence difference spectrum" /> |
− | <caption>Fig. - YFP emission spectrum of a <i>C. reinhardtii</i> clone with YFP with an emission maximum at approximately 530 nm. Difference spectrum of WT and YFP spectra</caption> | + | <caption>Fig. - YFP emission spectrum of a C. reinhardtii clone with YFP with an emission maximum at approximately 530 nm. Difference spectrum of WT and YFP spectra</caption> |
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| <img class="is-revealing" src="https://static.igem.org/mediawiki/2019/e/e5/T--Humboldt_Berlin--Psad-YFP-syn%28FL_OD%29-1day.png" alt="fluorescence intensity time resolved" /> | | <img class="is-revealing" src="https://static.igem.org/mediawiki/2019/e/e5/T--Humboldt_Berlin--Psad-YFP-syn%28FL_OD%29-1day.png" alt="fluorescence intensity time resolved" /> |