Part:BBa_K1017811:Experience

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Applications of BBa_K1017811

We used the following biobrick to test the efficiency of different RBSs:

P_cons+BBa_B0034+mRFP+Ter
P_cons+BBa_K1017202+mRFP+Ter
P_cons+BBa_B0030+mRFP+Ter
P_cons+BBa_B0032+mRFP+Ter
control: pet 30

As you can see from Figure 1 and Figure 2, the bacterial pellet of each biobrick shows different level of RFP expressions as each RBS provides a different translation efficiency. The deeper the red color is, the higher the level of expression is.

Figure 1. B0034 clearly expressed more RFP compared to others.

Figure 2. From left to right, the ribosome biding sites respectively: B0032, K1017202, B0030, B0034, control.

Figure 3. From left to right, the ribosome biding sites respectively: B0032, K1017202, B0030, B0034, control.

We measured the OD₆₀₀ value and the florescence expression (emmision 612 nm and excitation 584 nm) for each biobrick mentioned above. The result is shown in Figure 4. We calculated the normalized expression by dividing fluorescence expression with the OD value measured, since the higher the OD value is, the larger the amount of bacteria that can express florescence would be. As shown in Figure 4, the normalized expression of the biobrick with B0034 is the highest and the one with K1017202, is the second highest, while the other two of B0032 and B0030 show weak expressions. This result implies that K1017202 can, in fact, serve as a functional RBS. In comparison to other RBS, K1017202 can provide moderate translational efficiency that is just lower than that of the highly efficient B0034.

Figure 4. K1017202 shows moderate expression compared to the others.

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