Difference between revisions of "Part:BBa K1139150"
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[[Image:titech2013_parts_K1139150_main_Fig1.jpg|thumb|center|250px|<b>Fig. 1.</b> Our new designed hybrid promoter]]
 
[[Image:titech2013_parts_K1139150_main_Fig1.jpg|thumb|center|250px|<b>Fig. 1.</b> Our new designed hybrid promoter]]
  
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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 depending on the four different combinations of two inducers, CI and IPTG (Fig. 2). <br>  
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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 depending on the four different combinations of two inducers, CI and IPTG* (Fig. 2). <br>  
 
We confirmed that our new ''rm/lac'' hybrid promoter was actually activated by CI through an induction assay (Fig. 3).<br>
 
We confirmed that our new ''rm/lac'' hybrid promoter was actually activated by CI through an induction assay (Fig. 3).<br>
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<p>*We added IPTG in order to make sure to repress the expression of LacI derived from <i>E. coli</i> genome.
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</p>
  
 
[[Image:titech2013_parts_K1139150_main_Fig2.jpg|thumb|left|310px|<b>Fig. 2.</b> Fluorescence intensity detected by flow cytometer]]
 
[[Image:titech2013_parts_K1139150_main_Fig2.jpg|thumb|left|310px|<b>Fig. 2.</b> Fluorescence intensity detected by flow cytometer]]

Revision as of 02:19, 28 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 (Fig. 1).
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 depending on the four different combinations of two inducers, CI and IPTG* (Fig. 2).
We confirmed that our new rm/lac hybrid promoter was actually activated by CI through an induction assay (Fig. 3).

*We added IPTG in order to make sure to repress the expression of LacI derived from E. coli genome.

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