Regulatory
Prm/lac

Part:BBa_K1139151

Designed by: Naoki Watarai   Group: iGEM13_Tokyo_Tech   (2013-09-24)
Revision as of 02:19, 28 September 2013 by Fujimon (Talk | contribs)

rm/lac hybrid promoter

We newly developed rm/lac hybrid promoter which is activated by CI and repressed by LacI (Fig. 1).
To characterize the function of rm/lac hybrid promoter, we constructed a part Prm/lac-GFP (BBa_K1139150).

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. 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
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
n/arm/lac hybrid promoter