Difference between revisions of "Part:BBa K1139151"
 
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<partinfo>BBa_K1139151 short</partinfo>
 
<partinfo>BBa_K1139151 short</partinfo>
  
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We newly developed <i>rm/lac</i> hybrid promoter which is activated by CI and repressed by LacI (Fig. 1). <br>
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We newly developed the <i>RM/lac</i> hybrid promoter (<partinfo>BBa_K1139151</partinfo>) which is activated by CI and repressed by LacI (Fig. 1). <br>
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To characterize the function of <i>rm/lac</i> hybrid promoter, we constructed a part Prm/lac-GFP (<partinfo>BBa_K1139150</partinfo>).
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To characterize the function of this <i>RM/lac</i> hybrid promoter, we constructed a part PRM/lac-GFP (<partinfo>BBa_K1139150</partinfo>) by inserting the <i>RM/lac</i> promoter upstream of a GFP coding sequence.
  
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[[Image:titech2013_parts_K1139150_main_Fig1.jpg|thumb|center|250px|<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 newly 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 <i>rm/lac</i> 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. 2). We confirmed that our new <i>rm/lac</i> hybrid promoter was actually activated by CI through an induction assay (Fig. 3).
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By using the reporter cells that contain PRM/lac-GFP, we measured the fluorescence intensity of the cells induced by CI and IPTG (Fig. 2).  
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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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We saw that our new <i>RM/lac</i> hybrid promoter was actually activated by CI through an induction assay (Fig. 3).
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[[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]]

Latest revision as of 21:49, 28 October 2013

rm/lac hybrid promoter

We newly developed the RM/lac hybrid promoter (BBa_K1139151) which is activated by CI and repressed by LacI (Fig. 1).
To characterize the function of this RM/lac hybrid promoter, we constructed a part PRM/lac-GFP (BBa_K1139150) by inserting the RM/lac promoter upstream of a GFP coding sequence.

Fig. 1. Our newly designed hybrid promoter

By using the reporter cells that contain PRM/lac-GFP, we measured the fluorescence intensity of the cells induced by CI and IPTG (Fig. 2). We saw that our new RM/lac hybrid promoter was actually activated by CI through an induction assay (Fig. 3).

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
    COMPATIBLE WITH RFC[1000]