Difference between revisions of "Part:BBa K1139150"
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On the downstream of the promoter, GFP is inserted as a reporter.<br>
 
On the downstream of the promoter, GFP is inserted as a reporter.<br>
  
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[[Image:titech2013_parts_K1139150_main_Fig1.jpg|thumb|center|500px|<b>Fig. 1.</b> Our new designed hybrid promoter]]
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[[Image:titech2013_parts_K1139150_main_Fig1.jpg|thumb|center|250px|<b>Fig. 1.</b> Our new designed hybrid promoter]]
  
 
To characterize the function of the <i>rm/lac</i> hybrid promoter (<partinfo>BBa_K1139151</partinfo>), we constructed this part Prm/lac-GFP (<partinfo>BBa_K1139150</partinfo>) by inserting ''rm/lac'' promoter in front of a GFP coding sequence. By using the reporter cell that contains Prm/lac-GFP, we measured the fluorescence intensity of the reporter cell dependent on the four different combinations of two inducers, CI and IPTG (Fig. 1-B). <br>  
 
To characterize the function of the <i>rm/lac</i> hybrid promoter (<partinfo>BBa_K1139151</partinfo>), we constructed this part Prm/lac-GFP (<partinfo>BBa_K1139150</partinfo>) by inserting ''rm/lac'' promoter in front of a GFP coding sequence. By using the reporter cell that contains Prm/lac-GFP, we measured the fluorescence intensity of the reporter cell dependent on the four different combinations of two inducers, CI and IPTG (Fig. 1-B). <br>  

Revision as of 04:44, 27 September 2013

Prm/lac-GFP-TT

Prm/lac is a hybrid promoter that is modified to be activated by lamda repressor (CI) and repressed by LacI repressor.
On the downstream of the promoter, GFP is inserted as a reporter.

Fig. 1. Our new designed hybrid promoter

To characterize the function of the rm/lac hybrid promoter (BBa_K1139151), we constructed this part Prm/lac-GFP (BBa_K1139150) by inserting rm/lac promoter in front of a GFP coding sequence. By using the reporter cell that contains Prm/lac-GFP, we measured the fluorescence intensity of the reporter cell dependent on the four different combinations of two inducers, CI and IPTG (Fig. 1-B).
We confirmed that our new rm/lac hybrid promoter was actually activated by CI through an induction assay (Fig. 1-C).

Fig. 2. Fluorescence intensity detected by flow cytometer
Fig. 3. Comparison of N99 and JM2.300

For more information, see [http://2013.igem.org/Team:Tokyo_Tech/Experiment/RM-lac_Hybrid_Promoter_Assay our work in Tokyo_Tech 2013 wiki].

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 769