Difference between revisions of "Part:BBa K741003"

Latest revision as of 19:37, 2 October 2012

pRM-RBS-GFP-T

Promoter pRM with GFP gene downstream. The pRM is a modified one, whose OR3 (the third right operator) is mutated to be obliterated. This means our pRM is relatively difficult to be repressed by Cro dimer and can always be activated by CI dimer. We use this part to test the basic level of expression of modified pRM.

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 752

Figure 2 shows that the fluorescence intensity of prm-gfp is so weak in comparison with plac-gfp that we can assume that the leak expression of prm is very few, which doesn't affect our experiments.

Data

pRM-RBS-GFP-T
OD
t/min	Parallel	Experimental group	M9 medium
0	1	0.0593	0.045
	2	0.0642
	3	0.0678
45	1	0.0605	0.0436
	2	0.0734
	3	0.062
90	1	0.0648	0.0435
	2	0.0735
	3	0.0702
135	1	0.0811	0.0447
	2	0.0716
	3	0.0677
180	1	0.0723	0.0446
	2	0.0828
	3	0.0789

pRM-RBS-GFP-T
fluorescence intensity
t/min	Parallel	Experimental group	M9 medium
0	1	19.55	11.801
	2	19.571
	3	19.352
45	1	21.548	13.386
	2	20.505
	3	19.107
90	1	24.441	11.541
	2	23.777
	3	24.503
135	1	28.938	11.894
	2	27.884
	3	27.81
180	1	41.376	11.066
	2	39.573
	3	39.795

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K741003 short</partinfo>
-Express GFP only if protein CI activated the promoter pRM.
+Promoter pRM with GFP gene downstream. The pRM is a modified one, whose OR3 (the third right operator) is mutated to be obliterated.
+This means our pRM is relatively difficult to be repressed by Cro dimer and can always be activated by CI dimer. We use this part to test the basic level of expression of modified pRM.
 <!-- Add more about the biology of this part here
@@ Line 21: / Line 25: @@
   <tr height=19 style='height:14.25pt'>
    <td colspan=4 height=19 class=xl26 width=386 style='height:14.25pt;
-   width:290pt'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;pRM-RBS-GFP-T</td>
+   width:290pt'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;pRM-RBS-GFP-T</td>
   </tr>
   <tr height=19 style='height:14.25pt'>
-   <td colspan=4 height=19 class=xl26 style='height:14.25pt'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;OD</td>
+   <td colspan=4 height=19 class=xl26 style='height:14.25pt'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;OD</td>
   </tr>
   <tr height=19 style='height:14.25pt'>
@@ Line 144: / Line 148: @@
   </tr>
   <tr height=19 style='height:14.25pt'>
-   <td colspan=4 height=19 class=xl26 style='height:14.25pt'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;pRM-RBS-GFP-T</td>
+   <td colspan=4 height=19 class=xl26 style='height:14.25pt'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;pRM-RBS-GFP-T</td>
   </tr>
   <tr height=19 style='height:14.25pt'>
-   <td colspan=4 height=19 class=xl26 style='height:14.25pt'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;fluorescence
+   <td colspan=4 height=19 class=xl26 style='height:14.25pt'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;fluorescence
    intensity</td>
   </tr>