Difference between revisions of "Part:BBa K1139150"

Revision as of 03:31, 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 (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).

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

@@ Line 7: / Line 7: @@
 [[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).
+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).
 [[Image:Titech2013_parts_K1139150_Fig1B.jpg|thumb|center|500px|'''Fig. 1B.''' Fluorescence intensity detected by flow cytometer]]