Difference between revisions of "Part:BBa K2943902"
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<partinfo>BBa_K2943902 short</partinfo> | <partinfo>BBa_K2943902 short</partinfo> | ||
− | This part has a native promoter, an | + | This part is an improvemt of an existing composite part: INSERT PART.<br> |
+ | It has a native constitutive promoter, and contains improvement of Shine-Dalgarno(SD) sequence(including inside an improved RBS) and improvement of the monomer RFP (mRFP). The goal of the improvement was to enable more efficient and stronger expression of RFP through improving both the mRNA folding energy and through improvement of the SD that has direct impact on translation initiation and speed.<br> | ||
+ | <strong>The Experiment:</strong><br> | ||
+ | In order to check if our improvement worked we have designed an fluorescence experiment to compare between the original composite part and our version. Using 96 plate reader, we measured each part fluorescence intensity. In addition we also added wells for negative control (Bacteria with non fluorescence plasmid) and LB for blank. We have repeated the the measuring a few times through the day to make sure we do not get any false results and to make sure the bacteria had enough time to grow and express the protein. | ||
+ | The bacteria concentration was measured in OD700 to avoid any false measuring due to the absorption of light by the mRFP. Fluoresence was measured using the data below: | ||
+ | |||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here |
Revision as of 11:48, 19 October 2019
Promoter, RBS, mRFP (Improved)
This part is an improvemt of an existing composite part: INSERT PART.
It has a native constitutive promoter, and contains improvement of Shine-Dalgarno(SD) sequence(including inside an improved RBS) and improvement of the monomer RFP (mRFP). The goal of the improvement was to enable more efficient and stronger expression of RFP through improving both the mRNA folding energy and through improvement of the SD that has direct impact on translation initiation and speed.
The Experiment:
In order to check if our improvement worked we have designed an fluorescence experiment to compare between the original composite part and our version. Using 96 plate reader, we measured each part fluorescence intensity. In addition we also added wells for negative control (Bacteria with non fluorescence plasmid) and LB for blank. We have repeated the the measuring a few times through the day to make sure we do not get any false results and to make sure the bacteria had enough time to grow and express the protein.
The bacteria concentration was measured in OD700 to avoid any false measuring due to the absorption of light by the mRFP. Fluoresence was measured using the data below:
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 617
Illegal AgeI site found at 729 - 1000COMPATIBLE WITH RFC[1000]