Part:BBa_K593011:Experience

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Applications of BBa_K593011

For characterization of RNA thermometer, this composite was assembled without lysis casette (with promoter, ROSE and GFP). Two control groups were planned; first one is T7 promoter and GFP ligate, this is designed to measure expression level of T7 promoter by reading the GFP expression at 37 °C. The expected result was to observe the similar readings because without RNA thermometer temperature switch off device express similar readings at 37°C with RNA thermometer device at 42 °C. In the data analysis we observed that there is deviation in readings between control T7 with GFP at 37°C and to construct, T7 RNA thermometer and GFP at 42 °C. The second one is T7 promoter, ROSE and GFP ligate, this is designed to measure the expression level of T7 promoter with ROSE ligate at 37°C. This two groups were compared with T7 promoter ROSE and GFP ligate at 42 °C. The dimer formation of RNA thermometer at lower than 42 °C caused inefficient binding of RBS to DNA. That is, we measured the spectrophotometer and fluoresence spectrometer readings from protein formation to folding range.The GFP readings at two temperatures; however, at 42 °C the expression was observed in higher level. As seen at OD:0.6 reading there is a dramatic difference in expressions.

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