Difference between revisions of "Part:BBa K313000"
 
(One intermediate revision by the same user not shown)
Line 2: Line 2:
 
<partinfo>BBa_K313000 short</partinfo>
 
<partinfo>BBa_K313000 short</partinfo>
  
−
gfp generator with T7 promoter
+
gfp generator under the control of T7 promoter.
  
 
===Usage and Biology===
 
===Usage and Biology===
−
T7 RNA polymerase induces the expression of gfp from this part.
+
T7 RNA polymerase induces the expression of gfp from this part. This part can be used as reporter to monitor the expression of T7 polymerase.
  
 
=== Verification Experiment ===
 
=== Verification Experiment ===
−
Please see [http://2010.igem.org/Team:UT-Tokyo/Sudoku_assay_LeakSw  Terminator leak] assay page for detail.
+
(Please see [http://2010.igem.org/Team:UT-Tokyo/Sudoku_assay_LeakSw  Terminator leak] assay page for detail.)
  
−
Plasmid containing this part was transformed to Rosetta (DE3) plysS strain which have T7 polymerase under the control of Lac promoter.
+
Plasmid containing this part was transformed into Rosetta (DE3) plysS strain, which have T7 polymerase under the control of Lac promoter.
−
When the OD reached 0.6, we added IPTG to a final concentration of 0.1 mM
+
 
 +
When the OD reached 0.6, we added IPTG to a final concentration of 0.1 mM.
 
Fluorescence was measured and the results are shown below:
 
Fluorescence was measured and the results are shown below:
  
 
[[Image:Graph_5.png]]
 
[[Image:Graph_5.png]]
  
 +
Although we observed fluorescence without IPTG induction, the fluorescence intensity was half compared to that of IPTG induced sample.
 +
 +
As a result we successfully confirmed the proper operation of this part.
  
 
<!-- -->
 
<!-- -->

Latest revision as of 09:52, 3 November 2010

gfp generator with T7 promoter

gfp generator under the control of T7 promoter.

Usage and Biology

T7 RNA polymerase induces the expression of gfp from this part. This part can be used as reporter to monitor the expression of T7 polymerase.

Verification Experiment

(Please see [http://2010.igem.org/Team:UT-Tokyo/Sudoku_assay_LeakSw Terminator leak] assay page for detail.)

Plasmid containing this part was transformed into Rosetta (DE3) plysS strain, which have T7 polymerase under the control of Lac promoter.

When the OD reached 0.6, we added IPTG to a final concentration of 0.1 mM. Fluorescence was measured and the results are shown below:

Graph 5.png

Although we observed fluorescence without IPTG induction, the fluorescence intensity was half compared to that of IPTG induced sample.

As a result we successfully confirmed the proper operation of this part.

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 719