Difference between revisions of "Part:BBa K1151036"

(→‎Fluorescence decay assay)
(→‎Fluorescence decay assay)
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''The experiment''
 
''The experiment''
  
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We have set up this experiment to evaluate the shutdown of the fluorescence signal, adding a fixed quantity of IPTG and nickel to BL21 cells transformed with the plasmid containing this part. Then, we exploited the ability of Olo-NikR to bind the promoter pnik and repress the transcription of GFP (Nickel added 0.3 ul of stock 10 ug / ul; IPTG: 5 ul 1M).
+
We have set up this experiment to evaluate the shutdown of the fluorescence signal (using fluorimetry technique), adding a fixed quantity of IPTG and nickel to BL21 cells transformed with the plasmid containing this part. Then, we exploited the ability of Olo-NikR to bind the promoter pnik and repress the transcription of GFP (Nickel added 0.3 ul of stock 10 ug / ul; IPTG: 5 ul 1M).
 +
 
 +
''Results''
  
 
[[File:1036.2.jpg]]                          [[File:1036.3.jpg]]
 
[[File:1036.2.jpg]]                          [[File:1036.3.jpg]]
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[[File:decayas.jpg]]                        [[File:gfp2.jpg]]
 
[[File:decayas.jpg]]                        [[File:gfp2.jpg]]
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''Discussion''
  
 
===Sequence and Features===
 
===Sequence and Features===

Revision as of 09:29, 1 October 2013

Double generator NikR-GFP, IPTG-nickel regulated

A simple construct composed of the parts K1151006 + K1151009.


Fluorescence decay assay

The experiment

We have set up this experiment to evaluate the shutdown of the fluorescence signal (using fluorimetry technique), adding a fixed quantity of IPTG and nickel to BL21 cells transformed with the plasmid containing this part. Then, we exploited the ability of Olo-NikR to bind the promoter pnik and repress the transcription of GFP (Nickel added 0.3 ul of stock 10 ug / ul; IPTG: 5 ul 1M).

Results

1036.2.jpg 1036.3.jpg


Decayas.jpg Gfp2.jpg

Discussion

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 818
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 818
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 818
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 818
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 818
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1616