Difference between revisions of "Part:BBa K1139150"

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[[Image:Titech2013_parts_K1139150_Fig1A.jpg|thumb|center|500px|'''Fig. 1-A.''' Our new designed hybrid promoter]]
 
[[Image:Titech2013_parts_K1139150_Fig1A.jpg|thumb|center|500px|'''Fig. 1-A.''' Our new designed hybrid promoter]]
  
To characterize the function of the ''rm/lac'' hybrid promoter ([https://parts.igem.org/Part:BBa_K1139151 BBa_K1139151]), we constructed this part Prm/lac-GFP ([https://parts.igem.org/Part:BBa_K113950 BBa_K113950]) 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. We confirmed that our new ''rm/lac'' hybrid promoter was actually activated by CI through an induction assay (Fig. 1-C).
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To characterize the function of the ''rm/lac'' hybrid promoter (<partinfo>BBa_K1139151</partinfo>), we constructed this part Prm/lac-GFP (<partinfo>BBa_K113950</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>
 +
We confirmed that our new ''rm/lac'' hybrid promoter was actually activated by CI through an induction assay (Fig. 1-C).
  
 
[[Image:Titech2013_parts_K1139150_Fig1B.jpg|thumb|center|500px|'''Fig. 1B.''' Fluorescence intensity detected by flow cytometer]]
 
[[Image:Titech2013_parts_K1139150_Fig1B.jpg|thumb|center|500px|'''Fig. 1B.''' Fluorescence intensity detected by flow cytometer]]

Revision as of 03:52, 26 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-A. Our new designed hybrid promoter

To characterize the function of the rm/lac hybrid promoter (BBa_K1139151), we constructed this part Prm/lac-GFP (No part name specified with partinfo tag.) 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. 1B. Fluorescence intensity detected by flow cytometer
Fig. 1C. Comparison of N99 and JM2.300


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