Difference between revisions of "Part:BBa K1139150"
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Prm/lac is a hybrid promoter that is modified to be activated by lamda repressor (CI) and repressed by LacI repressor. | 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, | + | On the downstream of the promoter, ''GFP'' is inserted as a reporter. |
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+ | [[Image:Titech2013_parts_K1139150_Fig1A.jpg|thumb|center|500px|'''Fig. 1-A.''' Our new designed hybrid promoter]] | ||
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+ | 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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+ | [[Image:Titech2013_parts_K1139150_Fig1B.jpg|thumb|center|500px|'''Fig. 1B.''' Fluorescence intensity detected by flow cytometer]] | ||
+ | [[Image:Titech2013_parts_K1139150_Fig1C.jpg|thumb|center|500px|'''Fig. 1C.''' Comparison of N99 and JM2.300]] | ||
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Revision as of 02:02, 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.
To characterize the function of the rm/lac hybrid promoter (BBa_K1139151), we constructed this part Prm/lac-GFP (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).
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 769