Difference between revisions of "Part:BBa K741003"

(zs)
 
(7 intermediate revisions by 2 users not shown)
Line 2: Line 2:
 
<partinfo>BBa_K741003 short</partinfo>
 
<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
 
<!-- Add more about the biology of this part here
Line 17: Line 21:
  
  
<table x:str border=1 cellpadding=0 cellspacing=0 width=386 style='border-collapse:
+
<table border=1>
collapse;table-layout:fixed;width:290pt'>
+
  
 
  <tr height=19 style='height:14.25pt'>
 
  <tr height=19 style='height:14.25pt'>
 
   <td colspan=4 height=19 class=xl26 width=386 style='height:14.25pt;
 
   <td colspan=4 height=19 class=xl26 width=386 style='height:14.25pt;
   width:290pt'>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>
 
  <tr height=19 style='height:14.25pt'>
 
  <tr height=19 style='height:14.25pt'>
   <td colspan=4 height=19 class=xl26 style='height:14.25pt'>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>
 
  <tr height=19 style='height:14.25pt'>
 
  <tr height=19 style='height:14.25pt'>
Line 145: Line 148:
 
  </tr>
 
  </tr>
 
  <tr height=19 style='height:14.25pt'>
 
  <tr height=19 style='height:14.25pt'>
   <td colspan=4 height=19 class=xl26 style='height:14.25pt'>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>
 
  <tr height=19 style='height:14.25pt'>
 
  <tr height=19 style='height:14.25pt'>
   <td colspan=4 height=19 class=xl26 style='height:14.25pt'>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>
 
   intensity</td>
 
  </tr>
 
  </tr>

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

USTC2012Result2 副本.png

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