Reporter

Part:BBa_K1151036

Designed by: Gian Piero Lazzari   Group: iGEM13_UniSalento_Lecce   (2013-09-23)
Revision as of 09:42, 1 October 2013 by LeleBiotec (Talk | contribs) (→‎Fluorescence decay assay)

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 pnikR 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


[edit]
Categories
Parameters
n/aDouble generator NikR-GFP, IPTG-nickel regulated