PMC:4307275 / 21074-22768 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/4307275","sourcedb":"PMC","sourceid":"4307275","source_url":"https://www.ncbi.nlm.nih.gov/pmc/4307275","text":"On FCA, the comparison of fluorescence intensities between sGFP/K-sGFP (the cytoplasm) and KR-sGFP (chloroplasts) showed that chloroplast localization of sGFP increased the average of fluorescence intensity (see Figure 2A). These results could lead to the assumption that the difference in fluorescence intensity might derive from the expression levels. However, the results of real-time PCR and western blot analysis showed incongruously that their expression levels were rather similar to each other (see Figure 4). It suggests that the difference in fluorescence intensity is not related to their molecular levels and might be caused by other factors. Typically, the fluorescence emission (e.g., intensity, spectral features, fluorescence lifetime) are mostly limited to a small number of cellular conditions, such as pH, redox potential or refractive index, temperature, prior illumination, and protein concentration [24] in each protoplast cell, which could affect the emission efficiency of fluorescence from the fluorophore of sGFP. As another important factor that should not be overlooked, we could also consider the self-quenching caused by aggregation of high concentrated fluorophores [25]. However, microscopic analysis using a hemocytometer showed that the fluorescence emission degrees of K-sGFP and KR-sGFP are not considerably different from each other on the image of microscopic analysis using the same exposure time (see Figure 1B and Figure 2C), suggesting that the difference of fluorescence intensity as much as 8-fold might stem from not only cellular conditions or self-quenching influencing the efficiency of fluorescence emission, but also from other unknown factors.","tracks":[{"project":"2_test","denotations":[{"id":"25561231-9759496-50799911","span":{"begin":922,"end":924},"obj":"9759496"},{"id":"T60122","span":{"begin":922,"end":924},"obj":"9759496"}],"attributes":[{"subj":"25561231-9759496-50799911","pred":"source","obj":"2_test"},{"subj":"T60122","pred":"source","obj":"2_test"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"2_test","color":"#93ec9d","default":true}]}]}}