Difference between revisions of "Part:BBa K1139151"

Revision as of 02:11, 28 September 2013

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]

@@ Line 7: / Line 7: @@
 [[Image:titech2013_parts_K1139150_main_Fig1.jpg|thumb|center|250px|<b>Fig. 1.</b> Our new designed hybrid promoter]]
-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).
+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).
+<p>*We added IPTG in order to make sure to repress the expression of LacI derived from <i>E. coli</i> genome.
+</p>
 [[Image:titech2013_parts_K1139150_main_Fig2.jpg|thumb|left|310px|<b>Fig. 2.</b> Fluorescence intensity detected by flow cytometer]]