Part:BBa_K1017202:Experinece

37°C RBS

Using biobrick, P_cons + 37°C RBS + mGFP+J61048, we tested the function of the 37°C RBS at room temperature (around 25°C) and at 37°C.

Figure 1. The biobrick for testing the function of 37°C RBS.

Figure 2. The differences between the level of GFP expressions can be easily observed under UV light.

Figure 3. The differences between the level of GFP expressions.

As shown in Figure 4 the normalized expression, obtained by dividing the florescence expression (emmision 612 nm and excitation 584 nm) with the OD₆₀₀ value, of GFP under 37°C is much higher than the expression under room temperature. Such result demonstrates the fact that 37°C RBS can effectively regulate gene expression by responding to temperature. The increased kinetic energy at 37°C is sufficient to cause the 37°C RBS to unfold and become available for ribosome binding. At room temperature, however, there isn't sufficient kinetic energy to unfold the hairpin structure and the structure is preserved. As a result, the translational efficiency is very low at room tempersture.

Figure 4. The normalized expression (florescence expression/ OD value) under 37°C is higher than the expression under room temperature by about 6 folds.

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