Difference between revisions of "Part:BBa K1017202:Experinece"

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====rRBS efficiency====
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<p>We used the following biobrick to test the translational efficiency of K1017202 (rRBS) compared to the efficiency of other RBSs:
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#P<sub>cons</sub> + '''BBa_B0034''' + mRFP + Ter
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#P<sub>cons</sub> + '''[https://parts.igem.org/wiki/index.php?title=Part:BBa_K1017202 BBa_K1017202]'''+mRFP+Ter
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#P<sub>cons</sub> + '''BBa_B0030''' + mRFP + Ter
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#P<sub>cons</sub> + '''BBa_B0032''' + mRFP + Ter
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#control: pet 30
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</p>
  
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<p> As you can see from Figure 5, Figure 6, and Figure 7, the bacterial pellet and liquid of each biobrick shows different level of 'RFP expressions as the RBS of each biobrick provides a different translation efficiency. The deeper the red color is, the higher the level of expression is. </p>
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[[File:NCTU_Test_RBS_pic.png|center|600px|Figure 5. B0034 clearly expressed more ''RFP'' compared to others.]]
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[[File:NCTU_Test_RBS_flo_pic.png|center|600px|Figure 6. K1017202 shows moderate RFP expression that can be distinguied from the expressions B0030 and B0032 ]]
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[[File:NCTU_Test_RBS_ependorf.png|center|600px|Figure 7. From left to right, the ribosome biding sites respectively: B0032, K1017202, B0030, B0034, control.]]
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<p> We measured the normalized expression for each biobrick mentioned above. The result is shown in Figure 8. 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 8, the normalized expression of the biobrick with B0034 is the highest and the one with  K1017202 (rRBS) 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. </p>
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[[File:NCTU_Test_functional_test_of_different_RBS.png|center|600px|Figure 8. K1017202 shows moderate expression compared to the others.]]

Latest revision as of 18:40, 27 September 2013

rRBS efficiency

We used the following biobrick to test the translational efficiency of K1017202 (rRBS) compared to the efficiency of other RBSs:

  1. Pcons + BBa_B0034 + mRFP + Ter
  2. Pcons + BBa_K1017202+mRFP+Ter
  3. Pcons + BBa_B0030 + mRFP + Ter
  4. Pcons + BBa_B0032 + mRFP + Ter
  5. control: pet 30

As you can see from Figure 5, Figure 6, and Figure 7, the bacterial pellet and liquid of each biobrick shows different level of 'RFP expressions as the RBS of each biobrick provides a different translation efficiency. The deeper the red color is, the higher the level of expression is.

Figure 5. B0034 clearly expressed more RFP compared to others.
Figure 6. K1017202 shows moderate RFP expression that can be distinguied from the expressions B0030 and B0032
Figure 7. From left to right, the ribosome biding sites respectively: B0032, K1017202, B0030, B0034, control.


We measured the normalized expression for each biobrick mentioned above. The result is shown in Figure 8. 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 8, the normalized expression of the biobrick with B0034 is the highest and the one with K1017202 (rRBS) 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 8. K1017202 shows moderate expression compared to the others.