International Journal of Modern Biochemistry

نویسنده

  • Ibrahim Sani
چکیده

Application of green fluorescent protein (GFP) as a molecular biosensor or reporter is limited by several factors especially its poor fluorescence at low pH conditions. There are conflicting data on this limitation, however, systematic studies to assess the importance of this factor for growing bacterial cultures are lacking. In this study, effect of low pH on GFP green and red fluorescence intensities was demonstrated in batch cultures of Escherichia coli SCC1. The bacterial cells that express GFP were grown and photoactivated with blue light which made them absorb green light and emit red. In order to assess the fluorescence intensity under different pH conditions, relative fluorescence (RF) (absolute fluorescence/OD650) and percentage red-fluorescent to green (R/G (%)) were determined aerobically and anaerobically. The green and red fluorescence intensities were found to be reduced in pH values 4.0 and 5.0. The GFP green fluorescence has very low intensity of less than 70 RFU at pH 5.0 and about 20 RFU at pH 4.0. The intensity was shown to be higher (up to 110 RFU) at pH 6 7 in the aerobic cultures. It was shown that GFP red fluorescence changes with time in different pH conditions both aerobically and anaerobically. The red fluorescence intensity has higher magnitude anaerobically compared to the aerobic cultures. Thus, at pH levels 4 5, the red and green fluorescence intensities were both found to be significantly reduced.

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تاریخ انتشار 2013